肺癌肿瘤抑制物1（TSLC1）抗人肝癌细胞HepG2的实验研究
|
摘要
目的：探讨TSLC1基因抑制人肝癌细胞HepG2的机理,为肝癌的基因治疗提供有效的靶点。
方法：应用基因重组技术构建了真核表达载体pReceiver-M29-TSLC 1重组质粒,采用脂质体Lipofectamine 2000介导重组载体转染到HepG2肝癌细胞中,筛选后建立稳定转染细胞株,采用MTT法检测细胞增值情况、流式细胞仪检测细胞周期、利用酶标仪检测caspase-3和caspase-8酶活性,RT-PCR方法检测DAL-1mRNA的表达,探讨TSLC1真核表达载体对肝癌HepG2细胞的作用及其抗肝癌机制的实验研究。
结果：(1)成功的构建了pReceiver-M29-TSLC1重组质粒。(2)重组载体转染到HepG2肝癌细胞后,TSLC1mRNA表达明显增多,TSLC1蛋白表达明显增高；(3)我们成功证实了TSLC1能抑制细胞生长、阻滞细胞周期、提高caspase-3、caspase-8酶活性以及DAL-1的mRNA表达。
结论：TSLC1基因的表达可能通过抑制细胞周期、诱导肿瘤细胞凋亡、提高caspase-3、caspase-8酶活性及DAL-1mRNA的表达等方式起到抑制人肝癌细胞HepG2的作用,这为TSLC1作为肝癌基因治疗的有效靶点提供实验基础及理论依据。
Hepatocellular carcirioma(HCC)is reported to be the sixth most common cancer,and over 626,000 people newly issued in patients each year in the world, and about 50% of the patients occurred in china。Moreover,the mortality of HCC is the third highest among all cancers,only behind lung and colon cancer. Since 1990s and is now the second highest among all cancers.Hepatocellular carcinoma deaths increased to the second place in China. Liver resection is best way to treatment.of HCC,and could be supplemented by radiotherapy and chemotherapy.But about 15% of patients only could benefited from liver resection. Hepatocellular carcinoma cell are not sensitive to radiotherapy and chemotherapy. Five years survival rate is low, only 3%-6%,in hepatocellular carcinoma patients. At present,for the minimally invasive treatment of patients with hepatocellularcarcinoma is TACE, EIT, MTC. electric Chemotherapy treatment, RFA treatment, Immunotherapy, Brachytherapy. The efficient still reached less than 20%, the prognosis remains poor.
So,we are realized that we should find new ways to treatment of liver cancer based on mechanism of liver cancer. Studies have shown that the pathogenesis of liver cancer related to a variety of oncogene activation and inactivation of tumor suppressor genes. Choosen the key genes of oncogene or tumor suppressor genes of liver cancer, is expected to reach the purpose of treatment of liver cancer through gene therapy.
TSLC1, a tumor suppressor gene few studies,1993, Satoh speculated that the human chromosome 11 that there is a tumor suppressor gene. After years of study, 2002, Murakami et al transfected normal long arm of chromosome 11 DNA fragments into the NSCLC cell line A549 by functional complementation method,and then the transfected cells Injectied into the backs of nude mice,observed the DNA fragment can inhibit the tumor formation and confirmed human chromosome 11q23.2 has a new tumor suppressor gene, due to significantly inhibit the growth of lung cancer cell lines, so named for the TSLC1 (tumor suppressor in lung cancer 1, TSLC1).
TSLC1 gene mapping to human chromosome 11q23.2, the total length is more than 300kb, a total of 12 exons. TSLC1 gene 5', CpG islands, CpG island about the size of 900bp, is located in exon 1 and intron 1 upstream. In which exon 8a,8b and 8c in brain, there are many types of splicing isoforms, normal epithelial cells express only the exon 8a. TSLC1 gene transcripts of 4.4kb or 1.6kbmRNA precursors, translation 442 amino acid residues of the transmembrane glycoprotein. TSLC1 protein structure contains the extracellular domain, a transmembrane region and cytoplasmic region; these three regions may be related to TSLC1 protein has an important function of the relationship.It is inactivated in 44% of NSCLC and 30-60% of various cancers,including liver, pancreatic, and prostate cancers, especially in those with invasion or metastasis. Inactivation occurs by two hits:through promoter methylation, and through loss of heterozygosity at the gene locus. Now that promoter methylation is the main reason. The later stage cancer, the higher the degree of malignancy, TSLC1 promoter methylation frequency of the higher.
A great deal of research about TSLC1 anti-tumor mechanism of action At home and abroad, and its anti-tumor mechanism of action includes the following aspects: (1)TSLC1 associates with an actin-binding protein, DAL-1, and members of the membrane-associated guanylate kinase homologue (MAGuK)group, providing a novel tumor suppressor cascade that is inactivated in tumors.(2) TSLC1 as necl-2 induced tumor cells were phagocytosis by NK cell.(3) TSLC1, as antigen presented cells to CD8+T cells inhibit the occurrence of cancer.(4) TSLC1 may be activating apoptosis factor caspase-3, and raise its expression, promoting tumor cell apoptosis.(5) TSCL1 can inhibit tumor cell from the early DNA synthesis (G0/G1) phase to the DNA synthesis phase (S phase). And then inhibit tumor cell growth.
The TSLC1 research focused on lung cancer, leukemia, breast cancer, prostate cancer, breast cancer, but the research of TSLC1 in liver disease is still rare. at home and abroad.Although many studies have discovered that the expression of TSLC1 downregulate in patient of hepatoma carcinoma. But the study on mechanism of TSLC1 effect HepG2 hepatoma cell is lacking. This experiment was successfully constructed the eukaryotic expression vector pReceiver-M29-TSLC1, and the vector was transfected into HepG2 hepatoma cells line for research the mechanism of anti-tumor of TSLC1.and provide the theoretical and experimental basis for gene therapy. In this study, the research results obtained are as follows.
1. The construction of eukaryotic expression vector pReceiver-M29-TSLC1
The full length of TSLC1 gene was amplificated using specific primers for RT-PCR reaction by Genetic engineering technology,and linked with eukaryotic vector pReceiver-M29, and transformed into E.coli JM109. We identified this recombinant vector through application of restriction enzyme digestion, PCR and sequencing and confirmed that the TSLC1 gene fragment was correctly inserted into this eukaryotic expression vector.The results show that the eukaryotic expression vector pReceiver-M29-TSLC1 was constructed successfully, and established the experimental basis for further study of the mechanism in the inhibition of tumor.
2. Establish a stable TSLC1 gene transfected HepG2 cell line.
Eukaryotic expression vector pReceiver-M29-TSLC1 transfected into HepG2 hepatoma cells mediated through Lipofectamine 2000 liposome.G418 selection to establish a stable transfected cell lines.We detected that pReceiver-M29-TSLC1 has been transfected into HepG2 hepatoma cells by RT-PCR, immunohistochemical and westernblot methods.The experiment provides a cytological basis about TSLC1 and investigates the inhibition mechanism of TSLC1 in hepatoma cells HepG2.
3. TSLC1 can inhibit cell proliferation of hepatoma cells HepG2.
We detected pReceiver-M29-TSLC1-HepG2 group, pReceiver-M29-HepG2 and normal cell proliferation in HepG2 group by MTT methods. From the 3rd day,pReceiver-M29-TSLC1-HepG2 cells group growed slowly and proliferation were inhibited in a row of 7 days.We compared the MTT values between TSLC1-HepG2 cells and normal HepG2 cell group and found the significantly different of the value(P<0.05). Results showed that TSLC1 could inhibit hepatoma cells HepG2
4. TSLC1 could inhibit hepatoma cells HepG2 DNA synthesis from the early (GO/Gl phase) to the DNA synthesis phase (S phase) transformation.
We cultured pReceiver-M29-TSLC1-HepG2 cells and pReceiver-M29-HepG2 cells 3 days,and detected the change of cell cycle by the flow cytometry.We found early DNA synthesis cells (GO/G1 phase) were significantly increased and the DNA synthesis phase (S phase) cells decreased significantly in pReceiver-M29-TSLC1-HepG2 cells(P<0.05).Results show that the TSLC1 can inhibit DNA synthesis of hepatoma cells HepG2 in interphase of cell cycle, and then inhibit the growth of hepatoma cells HepG2.
5. TSLC1 can increase the caspase-3 significantly
We cultured the pReceiver-M29-TSLC1-HepG2 cells and pReceiver-M29-HepG2 cells 3 days,then detected the content of caspase-3. We compare the pReceiver-M29-TSLC1-HepG2 cells with pReceiver-M29-HepG2 cells found that the content of caspase-3 increased significantly in pReceiver-M29-TSLC1-HepG2 cells. The results showed that TSLC1 can increase the content of caspase-3 and caspase-8, leading to tumor cells apoptosis,then inhibit the growth of hepatoma cells HepG2.
6. TSLC1 can increase the caspase-8 significantly
We cultured the pReceiver-M29-TSLC1-HepG2 cells and pReceiver-M29-HepG2 cells 3 days,then detected the content of caspase-8. We compare the pReceiver-M29-TSLC1-HepG2 cells with pReceiver-M29-HepG2 cells found that the content of caspase-8 increased significantly in pReceiver-M29-TSLC1-HepG2 cells. The results showed that TSLC1 can increase the content of caspase-8, leading to tumor cells apoptosis,then inhibit the growth of hepatoma cells HepG2.
7. TSLC1 can increase the expression of the DAL-1 mRNA
We cultured the pReceiver-M29-TSLC1-HepG2 cells and pReceiver-M29-HepG2 cells 3 days,then detected the expression of DAL-1 mRNA.We compare the pReceiver-M29-TSLC1-HepG2 cells with pReceiver-M29-HepG2 cells found that the increased significantly expression of DAL-1 mRNA in pReceiver-M29-TSLCl-HepG2 cells. The results showed that TSLC1 can increase the expression of DAL-1 mRNA leading to inhibit the growth of hepatoma cells HepG2.
方法：应用基因重组技术构建了真核表达载体pReceiver-M29-TSLC 1重组质粒,采用脂质体Lipofectamine 2000介导重组载体转染到HepG2肝癌细胞中,筛选后建立稳定转染细胞株,采用MTT法检测细胞增值情况、流式细胞仪检测细胞周期、利用酶标仪检测caspase-3和caspase-8酶活性,RT-PCR方法检测DAL-1mRNA的表达,探讨TSLC1真核表达载体对肝癌HepG2细胞的作用及其抗肝癌机制的实验研究。
结果：(1)成功的构建了pReceiver-M29-TSLC1重组质粒。(2)重组载体转染到HepG2肝癌细胞后,TSLC1mRNA表达明显增多,TSLC1蛋白表达明显增高；(3)我们成功证实了TSLC1能抑制细胞生长、阻滞细胞周期、提高caspase-3、caspase-8酶活性以及DAL-1的mRNA表达。
结论：TSLC1基因的表达可能通过抑制细胞周期、诱导肿瘤细胞凋亡、提高caspase-3、caspase-8酶活性及DAL-1mRNA的表达等方式起到抑制人肝癌细胞HepG2的作用,这为TSLC1作为肝癌基因治疗的有效靶点提供实验基础及理论依据。
Hepatocellular carcirioma(HCC)is reported to be the sixth most common cancer,and over 626,000 people newly issued in patients each year in the world, and about 50% of the patients occurred in china。Moreover,the mortality of HCC is the third highest among all cancers,only behind lung and colon cancer. Since 1990s and is now the second highest among all cancers.Hepatocellular carcinoma deaths increased to the second place in China. Liver resection is best way to treatment.of HCC,and could be supplemented by radiotherapy and chemotherapy.But about 15% of patients only could benefited from liver resection. Hepatocellular carcinoma cell are not sensitive to radiotherapy and chemotherapy. Five years survival rate is low, only 3%-6%,in hepatocellular carcinoma patients. At present,for the minimally invasive treatment of patients with hepatocellularcarcinoma is TACE, EIT, MTC. electric Chemotherapy treatment, RFA treatment, Immunotherapy, Brachytherapy. The efficient still reached less than 20%, the prognosis remains poor.
So,we are realized that we should find new ways to treatment of liver cancer based on mechanism of liver cancer. Studies have shown that the pathogenesis of liver cancer related to a variety of oncogene activation and inactivation of tumor suppressor genes. Choosen the key genes of oncogene or tumor suppressor genes of liver cancer, is expected to reach the purpose of treatment of liver cancer through gene therapy.
TSLC1, a tumor suppressor gene few studies,1993, Satoh speculated that the human chromosome 11 that there is a tumor suppressor gene. After years of study, 2002, Murakami et al transfected normal long arm of chromosome 11 DNA fragments into the NSCLC cell line A549 by functional complementation method,and then the transfected cells Injectied into the backs of nude mice,observed the DNA fragment can inhibit the tumor formation and confirmed human chromosome 11q23.2 has a new tumor suppressor gene, due to significantly inhibit the growth of lung cancer cell lines, so named for the TSLC1 (tumor suppressor in lung cancer 1, TSLC1).
TSLC1 gene mapping to human chromosome 11q23.2, the total length is more than 300kb, a total of 12 exons. TSLC1 gene 5', CpG islands, CpG island about the size of 900bp, is located in exon 1 and intron 1 upstream. In which exon 8a,8b and 8c in brain, there are many types of splicing isoforms, normal epithelial cells express only the exon 8a. TSLC1 gene transcripts of 4.4kb or 1.6kbmRNA precursors, translation 442 amino acid residues of the transmembrane glycoprotein. TSLC1 protein structure contains the extracellular domain, a transmembrane region and cytoplasmic region; these three regions may be related to TSLC1 protein has an important function of the relationship.It is inactivated in 44% of NSCLC and 30-60% of various cancers,including liver, pancreatic, and prostate cancers, especially in those with invasion or metastasis. Inactivation occurs by two hits:through promoter methylation, and through loss of heterozygosity at the gene locus. Now that promoter methylation is the main reason. The later stage cancer, the higher the degree of malignancy, TSLC1 promoter methylation frequency of the higher.
A great deal of research about TSLC1 anti-tumor mechanism of action At home and abroad, and its anti-tumor mechanism of action includes the following aspects: (1)TSLC1 associates with an actin-binding protein, DAL-1, and members of the membrane-associated guanylate kinase homologue (MAGuK)group, providing a novel tumor suppressor cascade that is inactivated in tumors.(2) TSLC1 as necl-2 induced tumor cells were phagocytosis by NK cell.(3) TSLC1, as antigen presented cells to CD8+T cells inhibit the occurrence of cancer.(4) TSLC1 may be activating apoptosis factor caspase-3, and raise its expression, promoting tumor cell apoptosis.(5) TSCL1 can inhibit tumor cell from the early DNA synthesis (G0/G1) phase to the DNA synthesis phase (S phase). And then inhibit tumor cell growth.
The TSLC1 research focused on lung cancer, leukemia, breast cancer, prostate cancer, breast cancer, but the research of TSLC1 in liver disease is still rare. at home and abroad.Although many studies have discovered that the expression of TSLC1 downregulate in patient of hepatoma carcinoma. But the study on mechanism of TSLC1 effect HepG2 hepatoma cell is lacking. This experiment was successfully constructed the eukaryotic expression vector pReceiver-M29-TSLC1, and the vector was transfected into HepG2 hepatoma cells line for research the mechanism of anti-tumor of TSLC1.and provide the theoretical and experimental basis for gene therapy. In this study, the research results obtained are as follows.
1. The construction of eukaryotic expression vector pReceiver-M29-TSLC1
The full length of TSLC1 gene was amplificated using specific primers for RT-PCR reaction by Genetic engineering technology,and linked with eukaryotic vector pReceiver-M29, and transformed into E.coli JM109. We identified this recombinant vector through application of restriction enzyme digestion, PCR and sequencing and confirmed that the TSLC1 gene fragment was correctly inserted into this eukaryotic expression vector.The results show that the eukaryotic expression vector pReceiver-M29-TSLC1 was constructed successfully, and established the experimental basis for further study of the mechanism in the inhibition of tumor.
2. Establish a stable TSLC1 gene transfected HepG2 cell line.
Eukaryotic expression vector pReceiver-M29-TSLC1 transfected into HepG2 hepatoma cells mediated through Lipofectamine 2000 liposome.G418 selection to establish a stable transfected cell lines.We detected that pReceiver-M29-TSLC1 has been transfected into HepG2 hepatoma cells by RT-PCR, immunohistochemical and westernblot methods.The experiment provides a cytological basis about TSLC1 and investigates the inhibition mechanism of TSLC1 in hepatoma cells HepG2.
3. TSLC1 can inhibit cell proliferation of hepatoma cells HepG2.
We detected pReceiver-M29-TSLC1-HepG2 group, pReceiver-M29-HepG2 and normal cell proliferation in HepG2 group by MTT methods. From the 3rd day,pReceiver-M29-TSLC1-HepG2 cells group growed slowly and proliferation were inhibited in a row of 7 days.We compared the MTT values between TSLC1-HepG2 cells and normal HepG2 cell group and found the significantly different of the value(P<0.05). Results showed that TSLC1 could inhibit hepatoma cells HepG2
4. TSLC1 could inhibit hepatoma cells HepG2 DNA synthesis from the early (GO/Gl phase) to the DNA synthesis phase (S phase) transformation.
We cultured pReceiver-M29-TSLC1-HepG2 cells and pReceiver-M29-HepG2 cells 3 days,and detected the change of cell cycle by the flow cytometry.We found early DNA synthesis cells (GO/G1 phase) were significantly increased and the DNA synthesis phase (S phase) cells decreased significantly in pReceiver-M29-TSLC1-HepG2 cells(P<0.05).Results show that the TSLC1 can inhibit DNA synthesis of hepatoma cells HepG2 in interphase of cell cycle, and then inhibit the growth of hepatoma cells HepG2.
5. TSLC1 can increase the caspase-3 significantly
We cultured the pReceiver-M29-TSLC1-HepG2 cells and pReceiver-M29-HepG2 cells 3 days,then detected the content of caspase-3. We compare the pReceiver-M29-TSLC1-HepG2 cells with pReceiver-M29-HepG2 cells found that the content of caspase-3 increased significantly in pReceiver-M29-TSLC1-HepG2 cells. The results showed that TSLC1 can increase the content of caspase-3 and caspase-8, leading to tumor cells apoptosis,then inhibit the growth of hepatoma cells HepG2.
6. TSLC1 can increase the caspase-8 significantly
We cultured the pReceiver-M29-TSLC1-HepG2 cells and pReceiver-M29-HepG2 cells 3 days,then detected the content of caspase-8. We compare the pReceiver-M29-TSLC1-HepG2 cells with pReceiver-M29-HepG2 cells found that the content of caspase-8 increased significantly in pReceiver-M29-TSLC1-HepG2 cells. The results showed that TSLC1 can increase the content of caspase-8, leading to tumor cells apoptosis,then inhibit the growth of hepatoma cells HepG2.
7. TSLC1 can increase the expression of the DAL-1 mRNA
We cultured the pReceiver-M29-TSLC1-HepG2 cells and pReceiver-M29-HepG2 cells 3 days,then detected the expression of DAL-1 mRNA.We compare the pReceiver-M29-TSLC1-HepG2 cells with pReceiver-M29-HepG2 cells found that the increased significantly expression of DAL-1 mRNA in pReceiver-M29-TSLCl-HepG2 cells. The results showed that TSLC1 can increase the expression of DAL-1 mRNA leading to inhibit the growth of hepatoma cells HepG2.
引文
[1]Parkin DM, Bray F, Ferlay J, Pisani P. Global cancer statistics,2002. CA Cancer [J]J Clin.2005.55(2):74-108.
[2]El-Serag HB, Rudolph KL. Hepatocellular carcinoma:epidemiology and molecular carcinogenesis [J]. Gastroenterology.2007.132(7):2557-76.
[3]张思维,陈万青,雷正龙,邹小农,赵平.中国肿瘤登记处2004年恶性肿瘤发病资料分析[J].中国肿瘤.2008.(11)：909-912.
[4]Portolani N, Baiocchi GL, Coniglio A, et al. Sequential multidisciplinary treatment of hepatocellular carcinoma:the role of surgery as rescue therapy for failure of percutaneous ablation therapies [J]. J Surg Oncol.2009.100(7):580-4.
[5]Shao Z, Lopez R, Shen B, Yang GS. Orthotopic liver transplantation as a rescue operation for recurrent hepatocellular carcinoma after partial hepatectomy[J]. World Gastroenterol.2008.14(27):4370-6.
[6]Neuhaus P, Jonas S, Bechstein WO. Hepatoma of the liver-resection or transplantation [J]. Langenbecks Arch Surg.2000.385(3):171-8.
[7]Riordan SM, Williams R. Preoperative investigation and indication for operation in hepatocellular carcinoma [J]. Hepatobiliary Pancreat Surg.1998.5(1):1-6.
[8]Ding YT, Sun XT, Xu QX. Non-bleeding technique in resection of hepatoma: report of 49 cases[J]. Hepatobiliary Pancreat Dis Int.2002.1(1):52-6.
[9]Lang H, Radtke A, Hindennach M, et al. Impact of virtual tumor resection and computer-assisted risk analysis on operation planning and intraoperative strategy in major hepatic resection [J]. Arch Surg.2005.140(7):629-38; discussion 638.
[10]Llovet JM, Bruix J, Gores GJ. Surgical resection versus transplantation for early hepatocellular carcinoma:clues for the best strategy [J]. Hepatology.2000.31(4): 1019-21.
[11]Xu J, Shen ZY, Chen XG, et al. A randomized controlled trial of Licartin for preventing hepatoma recurrence after liver transplantation [J]. Hepatology.2007. 45(2):269-76.
[12]Shiga T, Noguchi T, Nishimaki K, Kudo M. [New approach for unresectable primary liver cancer-intra-arterial combination chemotherapy during and after operation with local injection of ethanol] [J]. Gan To Kagaku Ryoho.1989.16(8 Pt 2):2765-8.
[13]Ohnuma N, Takahashi H, Maie M, Etoh T, Tanabe M, Kuriyama Y. [Delayed primary operation of solid tumors in children] [J]. Nippon Geka Gakkai Zasshi. 1988.89(11):1914-9.
[14]Wang CH, Chen CL, Cheng KW, et al. Bispectral index monitoring in healthy, cirrhotic, and end-stage liver disease patients undergoing hepatic operation[J]. Transplant Proc.2008.40(8):2489-91.
[15]Jeng KS, Yang FS, Chiang HJ, Ohta I. Repeat operation for nodular recurrent hepatocellular carcinoma within the cirrhotic liver remnant:a comparison with transcatheter arterial chemoembolization[J]. World Surg.1992.16(6):1188-91; discussion 1192.
[16]Shimamura T, Une Y, Nakajima Y, et al. [Advantages and disadvantages of the intraarterial chemotherapy using a reservoir as postoperative adjuvant therapy for hepatocellular carcinoma] [J]. Gan To Kagaku Ryoho.1995.22(11):1511-4.
[17]Hayashida K, Ooi J, Makiyama K, Hara K. [Percutaneous ethanol injection therapy for hepatocellular carcinoma in hepatic dome directly under right diaphragma-the trial of artificial ascites method] [J]. Nippon Shokakibyo Gakkai Zasshi.1995.92(10):1752-8.
[18]Usui H, Itai Y. Percutaneous ethanol injection therapy for hepatocellular carcinoma:validity of MR imaging in the evaluation of treatment effect. Radiat Med[J].1995.13(3):103-8.
[19]Watanabe Y, Sato M, Abe Y, et al. Laparoscopic microwave coagulo-necrotic therapy for hepatocellular carcinoma:a feasible study of an alternative option for poor-risk patients[J]. Laparoendosc Surg.1995.5(3):169-75.
[20]Sato M, Watanabe Y, Ueda S, et al. Microwave coagulation therapy for hepatocellular carcinoma[J]. Gastroenterology.1996.110(5):1507-14.
[21]D'Agostino HB, Solinas A. Percutaneous ablation therapy for hepatocellular carcinomas[J].AJR Am J Roentgenol.1995.164(5):1165-7.
[22]Quick AM, Lo SS, Mayr NA, Kim EY. Radiation therapy for intrahepatic malignancies[J]. Expert Rev Anticancer Ther.2009.9(10):1511-21.
[23]Hilgard P, Muller S, Hamami M, et al. [Selective internal radiotherapy (radioembolization) and radiation therapy for HCC-current status and perspectives] [J]. Z Gastroenterol.2009.47(1):37-54.
[24]Leung SW, Huang EY, Cheng YF, Lu SN. Conformal radiation therapy for hepatoma with portal vein thrombosis[J]. Br J Radiol.2000.73(869):550-2.
[25]Shiratori I, Suzuki Y, Oshiumi H, et al. Recombinant interleukin-12 and interleukin-18 antitumor therapy in a guinea-pig hepatoma cell implant model [J]. Cancer Sci.2007.98(12):1936-42.
[26]Tepetes KN, Tsamandas AC, Felekouras ES, Salakou SG, Karavias DD. Conversion of unresectable hepatoma to resectable. Report of a case and review of the literature [J]. Hepatogastroenterology.2000.47(34):1105-9.
[27]Takahashi S, Matsuura N, Kurokawa T, Takahashi Y, Miura T. Co-operation of the transcription factor hepatocyte nuclear factor-4 with Sp1 or Sp3 leads to transcriptional activation of the human haem oxygenase-1 gene promoter in a hepatoma cell line [J]. Biochem J.2002.367(Pt 3):641-52.
[28]Kubo S, Kinoshita H, Hirohashi K, et al. Patterns of and risk factors for recurrence after liver resection for well-differentiated hepatocellular carcinoma:a special reference to multicentric carcinogenesis after operation [J]. Hepatogastroenterology.1999.46(30):3212-5.
[29]Hsieh YJ, Liu RS, Hwu L, et al. Cre/loxP system controlled by specific promoter for radiation-mediated gene therapy of hepatoma [J]. Anticancer Res.2007. 27(3B):1571-9.
[30]Ido A, Nakata K, Kato Y, et al. Gene therapy for hepatoma cells using a retrovirus vector carrying herpes simplex virus thymidine kinase gene under the control of human alpha-fetoprotein gene promoter [J]. Cancer Res.1995.55(14): 3105-9.
[31]Cheng J, Leng X, Peng J. [Construction of a hepatoma-targeting vector of adeno-associated virus containing human alpha-fetoprotein promoter and wild p53 gene in gene therapy of liver cancer] [J]. Zhonghua Yi Xue Za Zhi.2000. 80(6):461-3.
[32]Tang YC, Li Y, Qian GX. Reduction of tumorigenicity of SMMC-7721 hepatoma cells by vascular endothelial growth factor antisense gene therapy [J]. World J Gastroenterol.2001.7(1):22-7.
[33]Iwai M, Harada Y, Ishii M, et al. Suicide gene therapy of human hepatoma and its peritonitis carcinomatosis by a vector of replicative-deficient herpes simplex virus [J]. Biochem Biophys Res Commun.2002.291(4):855-60.
[34]Wang KM, Xia AD, Chen SS. [Combination therapy of experimental murine hepatoma with mIL-12 gene and MHC I gene'mediated by liposome] [J]. Ai Zheng.2002.21(10):1041-6.
[35]Lai D, Weng S, Wang C, et al. Small antisense RNA to cyclin D1 generated by pre-tRNA splicing inhibits growth of human hepatoma cells [J]. FEBS Lett.2004. 576(3):481-6.
[36]Cai X, Zhou J, Lin J, Sun X, Xue X, Li C. Experimental studies on PNP suicide gene therapy of hepatoma [J]. J Huazhong Univ Sci Techno log Med Sci.2005. 25(2):178-81.
[37]Kaneko S, Hallenbeck P, Kotani T, et al. Adenovirus-mediated gene therapy of hepatocellular carcinoma using cancer-specific gene expression [J]. Cancer Res. 1995.55(22):5283-7.
[38]Ganslmayer M, Ocker M, Zopf S, et al. A quadruple therapy synergistically blocks proliferation and promotes apoptosis of hepatoma cells [J]. Oncol Rep. 2004.11(5):943-50.
[39]Li PY, Huang HJ, Zhang Q. [A study on the apoptosis of hepatoma cells synergistically induced by plasmid-mediated anti-angiogenesis and immunopotentiation therapy] [J]. Zhonghua Gan Zang Bing Za Zhi.2008.16(12): 909-12.
[40]Li PY, Lin JS, Feng ZH, et al. Combined gene therapy of endostatin and interleukin 12 with polyvinylpyrrolidone induces a potent antitumor effect on hepatoma [J]. World J Gastroenterol.2004.10(15):2195-200.
[41]Satoh H, Lamb PW, Dong JT, et al. Suppression of tumorigenicity of A549 lung adenocarcinoma cells by human chromosomes 3 and 11 introduced via microcell-mediated chromosome transfer [J]. Mol Carcinog.1993.7(3):157-64.
[42]Murakami Y, Nobukuni T, Tamura K, et al. Localization of tumor suppressor activity important in nonsmall cell lung carcinoma on chromosome 11q [J]. Proc Natl Acad Sci U S A.1998.95(14):8153-8.
[43]Murakami Y. Functional cloning of a tumor suppressor gene, TSLC1, in human non-small cell lung cancer [J]. Oncogene.2002.21(45):6936-48.
[44]Tsujiuchi T, Sugata E, Masaoka T, et al. Expression and DNA methylation patterns of Tslcl and Dal-1 genes in hepatocellular carcinomas induced by N-nitrosodiethylamine in rats [J]. Cancer Sci.2007.98(7):943-8.
[45]李文涛,白华龙,周闯,宋燕,冯留顺,张水军.原发性肝癌中抑癌基因TSLC1蛋白的表达及其临床意义[J].第四军医大学学报.2007.(23)：2141-2143.
[46]Iizuka M, Sugiyama Y, Shiraishi M, Jones C, Sekiya T. Allelic losses in human chromosome 11 in lung cancers [J]. Genes Chromosomes Cancer.1995.13(1):40-6.
[47]Gomyo H, Arai Y, Tanigami A, et al. A 2-Mb sequence-ready contig map and a novel immunoglobulin superfamily gene IGSF4 in the LOH region of chromosome Ilq23.2 [J]. Genomics.1999.62(2):139-46.
[48]Kuramochi M, Fukuhara H, Nobukuni T, et al. TSLC1 is a tumor-suppressor gene in human non-small-cell lung cancer [J]. Nat Genet.2001.27(4):427-30.
[49]Masuda M, Yageta M, Fukuhara H, et al. The tumor suppressor protein TSLC1 is involved in cell-cell adhesion [J]. Biol Chem.2002.277(34):31014-9.
[50]Ito A, Hagiyama M, Mimura T, et al. Expression of cell adhesion molecule 1 in malignant pleural mesothelioma as a cause of efficient adhesion and growth on mesothelium [J]. Lab Invest.2008.88(5):504-14.
[51]Koma Y, Furuno T, Hagiyama M, et al. Cell adhesion molecule 1 is a novel pancreatic-islet cell adhesion molecule that mediates nerve-islet cell interactions [J]. Gastroenterology.2008.134(5):1544-54.
[52]Fujita E, Soyama A, Momoi T. RA175, which is the mouse ortholog of TSLC1, a tumor suppressor gene in human lung cancer, is a cell adhesion molecule [J]. Exp Cell Res.2003.287(1):57-66.
[53]Watabe K, Ito A, Koma YI, Kitamura Y. IGSF4:a new intercellular adhesion molecule that is called by three names, TSLC1, SgIGSF and SynCAM, by virtue of its diverse function [J]. Histol Histopathol.2003.18(4):1321-9.
[54]Shingai T, Ikeda W, Kakunaga S, et al. Implications of nectin-like molecule-2/IGSF4/RA175/SgIGSF/TSLC1/SynCAMl in cell-cell adhesion and transmembrane protein localization in epithelial cells [J]. J Biol Chem.2003. 278(37):35421-7.
[55]Mandai K, Nakanishi H, Satoh A, et al. Afadin:A novel actin filament-binding protein with one PDZ domain localized at cadherin-based cell-to-cell adherens junction [J]. J Cell Biol.1997.139(2):517-28.
[56]Mandai K, Nakanishi H, Satoh A, et al. Ponsin/SH3P12:an 1-afadin-and vinculin-binding protein localized at cell-cell and cell-matrix adherens junctions [J]. J Cell Biol.1999.144(5):1001-17.
[57]Fukuhara H, Masuda M, Yageta M, et al. Association of a lung tumor suppressor TSLC1 with MPP3, a human homologue of Drosophila tumor suppressor Dlg [J]. Oncogene.2003.22(40):6160-5.
[58]Fukami T, Satoh H, Fujita E, et al. Identification of the Tslc1 gene, a mouse orthologue of the human tumor suppressor TSLC1 gene [J]. Gene.2002.295(1):7-12.
[59]Hoover KB, Bryant PJ. The genetics of the protein 4.1 family:organizers of the membrane and cytoskeleton [J]. Curr Opin Cell Biol.2000.12(2):229-34.
[60]Mangeat P, Roy C, Martin M. ERM proteins in cell adhesion and membrane dynamics [J]. Trends Cell Biol.1999.9(5):187-92.
[61]Tran YK, Bogler O, Gorse KM, Wieland I, Green MR, Newsham IF. A novel member of the NF2/ERM/4.1 superfamily with growth suppressing properties in lung cancer [J]. Cancer Res.1999.59(1):35-43.
[62]Galibert L, Diemer GS,Liu Z, et al. Nectin-like protein 2 defines a subset of T-cell zone dendritic cells and is a ligand for class-I-restricted T-cell-associated molecule [J]. J Biol Chem.2005.280(23):21955-64.
[63]Boles KS, Barchet W, Diacovo T, Cella M, Colonna M. The tumor suppressor TSLC1/NECL-2 triggers NK-cell and CD8+ T-cell responses through the cell-surface receptor CRTAM [J]. Blood.2005.106(3):779-86.
[64]Kakunaga S, Ikeda W, Itoh S, et al. Nectin-like molecule-1/TSLL1/SynCAM3:a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule localizing at non-junctional contact sites of presynaptic nerve terminals, axons and glia cell processes [J]. J Cell Sci.2005.118(Pt 6):1267-77.
[65]Fukuhara H, Kuramochi M, Nobukuni T, et al. Isolation of the TSLL1 and TSLL2 genes, members of the tumor suppressor TSLC1 gene family encoding transmembrane proteins [J]. Oncogene.2001.20(38):5401-7.
[66]Fujita E, Kouroku Y, Ozeki S, et al. Oligo-astheno-teratozoospermia in mice lacking RA175/TSLC1/SynCAM/IGSF4A, a cell adhesion molecule in the immunoglobulin superfamily [J]. Mol Cell Biol.2006.26(2):718-26.
[67]Wakayama T, Koami H, Ariga H, et al. Expression and functional characterization of the adhesion molecule spermatogenic immunoglobulin superfamily in the mouse testis [J]. Biol Reprod.2003.68(5):1755-63.
[68]Ito A, Jippo T, Wakayama T, et al. SgIGSF:a new mast-cell adhesion molecule used for attachment to fibroblasts and transcriptionally regulated by MITF [J]. Blood.2003.101(7):2601-8.
[69]Biederer T. Bioinformatic characterization of the SynCAM family of immunoglobulin-like domain-containing adhesion molecules [J]. Genomics. 2006.87(1):139-50.
[70]Hirohashi S, Kanai Y. Cell adhesion system and human cancer morphogenesis [J]. Cancer Sci.2003.94(7):575-81.
[71]Mao X, Seidlitz E, Ghosh K, Murakami Y, Ghosh HP. The cytoplasmic domain is critical to the tumor suppressor activity of TSLC1 in non-small cell lung cancer [J]. Cancer Res.2003.63(22):7979-85.
[72]Yageta M, Kuramochi M, Masuda M, et al. Direct association of TSLC1 and DAL-1, two distinct tumor suppressor proteins in lung cancer [J]. Cancer Res. 2002.62(18):5129-33.
[73]Apoptosis [J]. Lancet.1972.2(7785):1011-2.
[74]da FRR, Kosiol C, Vinar T, Siepel A, Nielsen R. Positive selection on apoptosis related genes [J]. FEBS Lett.2010.584(3):469-76.
[75]Pop C, Salvesen GS. Human caspases:activation, specificity, and regulation [J]. J Biol Chem.2009.284(33):21777-81.
[76]Kurokawa M, Kornbluth S. Caspases and kinases in a death grip [J]. Cell.2009. 138(5):838-54.
[77]Mao X, Seidlitz E, Truant R, Hitt M, Ghosh HP. Re-expression of TSLC1 in a non-small-cell lung cancer cell line induces apoptosis and inhibits tumor growth [J]. Oncogene.2004.23(33):5632-42.
[78]Sussan TE, Pletcher MT, Murakami Y, Reeves RH. Tumor suppressor in lung cancer 1 (TSLC1) alters tumorigenic growth properties and gene expression [J]. Mol Cancer.2005.4:28.
[79]Jiang W, Newsham IF. The tumor suppressor DAL-1/4.1B and protein methylation cooperate in inducing apoptosis in MCF-7 breast cancer cells [J]. Mol Cancer.2006.5:4.
[80]Grimm M, Gasser M, Bueter M, et al. Evaluation of immunological escape mechanisms in a mouse model of colorectal liver metastases [J]. BMC Cancer. 2010.10:82.
[81]Palena C, Schlom J. Vaccines against Human Carcinomas:Strategies to Improve Antitumor Immune Responses [J]. Biomed Biotechnol.2010.2010:380697.
[82]Sasaki H, Nishikata I, Shiraga T, et al. Overexpression of a cell adhesion molecule, TSLC1, as a possible molecular marker for acute-type adult T-cell leukemia [J]. Blood.2005.105(3):1204-13.
[83]Dewan MZ, Takamatsu N, Hidaka T, et al. Critical role for TSLC1 expression in the growth and organ infiltration of adult T-cell leukemia cells in vivo [J]. J Virol. 2008.82(23):11958-63.
[84]Kitamura Y, Kurosawa G, Tanaka M, et al. Frequent overexpression of CADM1/IGSF4 in lung adenocarcinoma [J]. Biochem Biophys Res Commun. 2009.383(4):480-4.
[85]Fukuhara H, Kuramochi M, Fukami T, et al. Promoter methylation of TSLC1 and tumor suppression by its gene product in human prostate cancer [J]. Jpn J Cancer Res.2002.93(6):605-9.
[86]Jansen M, Fukushima N, Rosty C, et al. Aberrant methylation of the 5'CpG island of TSLC1 is common in pancreatic ductal adenocarcinoma and is first manifest in high-grade PanlNs [J]. Cancer Biol Ther.2002.1(3):293-6.
[87]Allinen M, Peri L, Kujala S, et al. Analysis of 11q21-24 loss of heterozygosity candidate target genes in breast cancer:indications of TSLC1 promoter hypermethylation [J]. Genes Chromosomes Cancer.2002.34(4):384-9.
[88]Steenbergen RD, Kramer D, Braakhuis BJ, et al. TSLC1 gene silencing in cervical cancer cell lines and cervical neoplasia [J]. J Natl Cancer Inst.2004. 96(4):294-305.
[89]Overmeer RM, Henken FE, Snijders PJ, et al. Association between dense CADM1 promoter methylation and reduced protein expression in high-grade CIN and cervical SCC [J]. J Pathol.2008.215(4):388-97.
[90]Fu L, Gao Z, Zhang X, et al. Frequent concomitant epigenetic silencing of the stress-responsive tumor suppressor gene CADM1, and its interacting partner DAL-1 in nasal NK/T-cell lymphoma [J]. Int J Cancer.2009.124(7):1572-8.
[91]Honda T, Tamura G, Waki T, et al. Hypermethylation of the TSLC1 gene promoter in primary gastric cancers and gastric cancer cell lines [J]. Jpn J Cancer Res.2002.93(8):857-60.
[92]Nowacki S, Skowron M, Oberthuer A, et al. Expression of the tumour suppressor gene CADM1 is associated with favourable outcome and inhibits cell survival in neuroblastoma [J]. Oncogene.2008.27(23):3329-38.
[93]Ando K, Ohira M, Ozaki T, et al. Expression of TSLC1, a candidate tumor suppressor gene mapped to chromosome 11q23, is downregulated in unfavorable neuroblastoma without promoter hypermethylation [J]. Int J Cancer.2008. 123(9):2087-94.
[94]Kikuchi S, Yamada D, Fukami T, et al. Hypermethylation of the TSLC1/IGSF4 promoter is associated with tobacco smoking and a poor prognosis in primary nonsmall cell lung carcinoma [J]. Cancer.2006.106(8):1751-8.
[95]Goto A, Niki T, Chi-Pin L, et al. Loss of TSLC1 expression in lung adenocarcinoma:relationships with histological subtypes, sex and prognostic significance [J]. Cancer Sci.2005.96(8):480-6.
[96]Fukami T, Fukuhara H, Kuramochi M, et al. Promoter methylation of the TSLC1 gene in advanced lung tumors and various cancer cell lines [J]. Int J Cancer. 2003.107(1):53-9.
[97]Uchino K, Ito A, Wakayama T, et al. Clinical implication and prognostic significance of the tumor suppressor TSLC1 gene detected in adenocarcinoma of the lung [J]. Cancer.2003.98(5):1002-7.
[98]Ito T, Shimada Y, Hashimoto Y, et al. Involvement of TSLC1 in progression of esophageal squamous cell carcinoma [J]. Cancer Res.2003.63(19):6320-6.
[99]Surace El, Lusis E, Murakami Y, Scheithauer BW, Perry A, Gutmann DH. Loss of tumor suppressor in lung cancer-1 (TSLC1) expression in meningioma correlates with increased malignancy grade and reduced patient survival [J]. J Neuropathol Exp Neurol.2004.63(10):1015-27.
[100]Shimizu K, Onishi M, Sugata E, Fujii H, Honoki K, Tsujiuchi T. Aberrant DNA methylation of the 5'upstream region of Tslc1 gene in hamster pancreatic tumors [J]. Biochem Biophys Res Commun.2007.353(2):522-6.
[101]Borinstein SC, Conerly M, Dzieciatkowski S, et al. Aberrant DNA methylation occurs in colon neoplasms arising in the azoxymethane colon cancer model [J]. Mol Carcinog.2010.49(1):94-103.
[102]袁林,陈安民,郭风劲,祝文涛,王江,廖晖.肺癌抑癌基因1对人前列腺癌T_3B细胞增殖的抑制效应[J].中国癌症杂志.2009.(01)：1-5.
[103]Heller G, Geradts J, Ziegler B, et al. Downregulation of TSLC1 and DAL-1 expression occurs frequently in breast cancer [J]. Breast Cancer Res Treat. 2007.103(3)：283-91.
[104]丁勇,李小荣,杨开焰,胡桂.乳腺癌组织中肺癌肿瘤抑制物1的表达及意义[J].实用预防医学.2009.(02)：355-357.
[105]Yang YX, Yang AH, Yang ZJ, Wang ZR, Xia XH. Involvement of tumor suppressor in lung cancer 1 gene expression in cervical carcinogenesis [J]. Int J Gynecol Cancer.2006.16(5):1868-72.
[106]Usami Y, Ito A, Ohnuma K, Fuku T, Komori T, Yokozaki H. Tumor suppressor in lung cancer-1 as a novel ameloblast adhesion molecule and its downregulation in ameloblastoma [J]. Pathol Int.2007.57(2):68-75.
[107]Hori RT. The minimal Tumor Suppressor in Lung Cancer-1 promoter is restrained by an inhibitory region [J]. Molecular Biology Reports.2009.
[108]Bhatia R, Dogra RC, Sharma PK. Construction of green fluorescent protein (GFP)-marked strains of Bradyrhizobium for ecological studies [J]. J Appl Microbiol.2002.93(5):835-9.
[109]Koczor CA, Snyder JW, Shokolenko IN, Dobson AW, Wilson GL, Ledoux SP. Targeting repair proteins to the mitochondria of mammalian cells through stable transfection, transient transfection, viral transduction, and TAT-mediated protein transduction [J]. Methods Mol Biol.2009.554: 233-49.
[110]Wright JF. Transient transfection methods for clinical adeno-associated viral vector production [J]. Hum Gene Ther.2009.20(7):698-706.
[111]Townsend-Nicholson A. Approaches to the stable transfection of G protein-coupled receptors [J]. Methods Mol Biol.1997.83:45-54.
[112]Douglas KL. Toward development of artificial viruses for gene therapy:a comparative evaluation of viral and non-viral transfection [J]. Biotechnol Prog.2008.24(4):871-83.
[113]Montier T, Benvegnu T, Jaffres PA, Yaouanc JJ, Lehn P. Progress in cationic lipid-mediated gene transfection:a series of bio-inspired lipids as an example [J]. Curr Gene Ther.2008.8(5):296-312.
[114]Susa T, Kato T, Kato Y. Reproducible transfection in the presence of carrier DNA using FuGENE6 and Lipofectamine2000 [J]. Mol Biol Rep.2008. 35(3):313-9.
[115]章静波.细胞生物学实用方法与技术[M].北京医科大学一中国协和医科 大学联合出版社.1995：581-597.
[116]Yamada D, Yoshida M, Williams YN, et al. Disruption of spermatogenic cell adhesion and male infertility in mice lacking TSLC1/IGSF4, an immunoglobulin superfamily cell adhesion molecule [J]. Mol Cell Biol.2006. 26(9):3610-24.
[117]Heller G, Fong KM, Girard L, et al. Expression and methylation pattern of TSLC1 cascade genes in lung carcinomas [J]. Oncogene.2006.25(6): 959-68.
[118]覃莉,张正茂,郝友华等.肺癌抑癌基因1对HepG2细胞生长的抑制效应[J].中华肝脏病杂志.2007.15(7)：509-512.
[2]El-Serag HB, Rudolph KL. Hepatocellular carcinoma:epidemiology and molecular carcinogenesis [J]. Gastroenterology.2007.132(7):2557-76.
[3]张思维,陈万青,雷正龙,邹小农,赵平.中国肿瘤登记处2004年恶性肿瘤发病资料分析[J].中国肿瘤.2008.(11)：909-912.
[4]Portolani N, Baiocchi GL, Coniglio A, et al. Sequential multidisciplinary treatment of hepatocellular carcinoma:the role of surgery as rescue therapy for failure of percutaneous ablation therapies [J]. J Surg Oncol.2009.100(7):580-4.
[5]Shao Z, Lopez R, Shen B, Yang GS. Orthotopic liver transplantation as a rescue operation for recurrent hepatocellular carcinoma after partial hepatectomy[J]. World Gastroenterol.2008.14(27):4370-6.
[6]Neuhaus P, Jonas S, Bechstein WO. Hepatoma of the liver-resection or transplantation [J]. Langenbecks Arch Surg.2000.385(3):171-8.
[7]Riordan SM, Williams R. Preoperative investigation and indication for operation in hepatocellular carcinoma [J]. Hepatobiliary Pancreat Surg.1998.5(1):1-6.
[8]Ding YT, Sun XT, Xu QX. Non-bleeding technique in resection of hepatoma: report of 49 cases[J]. Hepatobiliary Pancreat Dis Int.2002.1(1):52-6.
[9]Lang H, Radtke A, Hindennach M, et al. Impact of virtual tumor resection and computer-assisted risk analysis on operation planning and intraoperative strategy in major hepatic resection [J]. Arch Surg.2005.140(7):629-38; discussion 638.
[10]Llovet JM, Bruix J, Gores GJ. Surgical resection versus transplantation for early hepatocellular carcinoma:clues for the best strategy [J]. Hepatology.2000.31(4): 1019-21.
[11]Xu J, Shen ZY, Chen XG, et al. A randomized controlled trial of Licartin for preventing hepatoma recurrence after liver transplantation [J]. Hepatology.2007. 45(2):269-76.
[12]Shiga T, Noguchi T, Nishimaki K, Kudo M. [New approach for unresectable primary liver cancer-intra-arterial combination chemotherapy during and after operation with local injection of ethanol] [J]. Gan To Kagaku Ryoho.1989.16(8 Pt 2):2765-8.
[13]Ohnuma N, Takahashi H, Maie M, Etoh T, Tanabe M, Kuriyama Y. [Delayed primary operation of solid tumors in children] [J]. Nippon Geka Gakkai Zasshi. 1988.89(11):1914-9.
[14]Wang CH, Chen CL, Cheng KW, et al. Bispectral index monitoring in healthy, cirrhotic, and end-stage liver disease patients undergoing hepatic operation[J]. Transplant Proc.2008.40(8):2489-91.
[15]Jeng KS, Yang FS, Chiang HJ, Ohta I. Repeat operation for nodular recurrent hepatocellular carcinoma within the cirrhotic liver remnant:a comparison with transcatheter arterial chemoembolization[J]. World Surg.1992.16(6):1188-91; discussion 1192.
[16]Shimamura T, Une Y, Nakajima Y, et al. [Advantages and disadvantages of the intraarterial chemotherapy using a reservoir as postoperative adjuvant therapy for hepatocellular carcinoma] [J]. Gan To Kagaku Ryoho.1995.22(11):1511-4.
[17]Hayashida K, Ooi J, Makiyama K, Hara K. [Percutaneous ethanol injection therapy for hepatocellular carcinoma in hepatic dome directly under right diaphragma-the trial of artificial ascites method] [J]. Nippon Shokakibyo Gakkai Zasshi.1995.92(10):1752-8.
[18]Usui H, Itai Y. Percutaneous ethanol injection therapy for hepatocellular carcinoma:validity of MR imaging in the evaluation of treatment effect. Radiat Med[J].1995.13(3):103-8.
[19]Watanabe Y, Sato M, Abe Y, et al. Laparoscopic microwave coagulo-necrotic therapy for hepatocellular carcinoma:a feasible study of an alternative option for poor-risk patients[J]. Laparoendosc Surg.1995.5(3):169-75.
[20]Sato M, Watanabe Y, Ueda S, et al. Microwave coagulation therapy for hepatocellular carcinoma[J]. Gastroenterology.1996.110(5):1507-14.
[21]D'Agostino HB, Solinas A. Percutaneous ablation therapy for hepatocellular carcinomas[J].AJR Am J Roentgenol.1995.164(5):1165-7.
[22]Quick AM, Lo SS, Mayr NA, Kim EY. Radiation therapy for intrahepatic malignancies[J]. Expert Rev Anticancer Ther.2009.9(10):1511-21.
[23]Hilgard P, Muller S, Hamami M, et al. [Selective internal radiotherapy (radioembolization) and radiation therapy for HCC-current status and perspectives] [J]. Z Gastroenterol.2009.47(1):37-54.
[24]Leung SW, Huang EY, Cheng YF, Lu SN. Conformal radiation therapy for hepatoma with portal vein thrombosis[J]. Br J Radiol.2000.73(869):550-2.
[25]Shiratori I, Suzuki Y, Oshiumi H, et al. Recombinant interleukin-12 and interleukin-18 antitumor therapy in a guinea-pig hepatoma cell implant model [J]. Cancer Sci.2007.98(12):1936-42.
[26]Tepetes KN, Tsamandas AC, Felekouras ES, Salakou SG, Karavias DD. Conversion of unresectable hepatoma to resectable. Report of a case and review of the literature [J]. Hepatogastroenterology.2000.47(34):1105-9.
[27]Takahashi S, Matsuura N, Kurokawa T, Takahashi Y, Miura T. Co-operation of the transcription factor hepatocyte nuclear factor-4 with Sp1 or Sp3 leads to transcriptional activation of the human haem oxygenase-1 gene promoter in a hepatoma cell line [J]. Biochem J.2002.367(Pt 3):641-52.
[28]Kubo S, Kinoshita H, Hirohashi K, et al. Patterns of and risk factors for recurrence after liver resection for well-differentiated hepatocellular carcinoma:a special reference to multicentric carcinogenesis after operation [J]. Hepatogastroenterology.1999.46(30):3212-5.
[29]Hsieh YJ, Liu RS, Hwu L, et al. Cre/loxP system controlled by specific promoter for radiation-mediated gene therapy of hepatoma [J]. Anticancer Res.2007. 27(3B):1571-9.
[30]Ido A, Nakata K, Kato Y, et al. Gene therapy for hepatoma cells using a retrovirus vector carrying herpes simplex virus thymidine kinase gene under the control of human alpha-fetoprotein gene promoter [J]. Cancer Res.1995.55(14): 3105-9.
[31]Cheng J, Leng X, Peng J. [Construction of a hepatoma-targeting vector of adeno-associated virus containing human alpha-fetoprotein promoter and wild p53 gene in gene therapy of liver cancer] [J]. Zhonghua Yi Xue Za Zhi.2000. 80(6):461-3.
[32]Tang YC, Li Y, Qian GX. Reduction of tumorigenicity of SMMC-7721 hepatoma cells by vascular endothelial growth factor antisense gene therapy [J]. World J Gastroenterol.2001.7(1):22-7.
[33]Iwai M, Harada Y, Ishii M, et al. Suicide gene therapy of human hepatoma and its peritonitis carcinomatosis by a vector of replicative-deficient herpes simplex virus [J]. Biochem Biophys Res Commun.2002.291(4):855-60.
[34]Wang KM, Xia AD, Chen SS. [Combination therapy of experimental murine hepatoma with mIL-12 gene and MHC I gene'mediated by liposome] [J]. Ai Zheng.2002.21(10):1041-6.
[35]Lai D, Weng S, Wang C, et al. Small antisense RNA to cyclin D1 generated by pre-tRNA splicing inhibits growth of human hepatoma cells [J]. FEBS Lett.2004. 576(3):481-6.
[36]Cai X, Zhou J, Lin J, Sun X, Xue X, Li C. Experimental studies on PNP suicide gene therapy of hepatoma [J]. J Huazhong Univ Sci Techno log Med Sci.2005. 25(2):178-81.
[37]Kaneko S, Hallenbeck P, Kotani T, et al. Adenovirus-mediated gene therapy of hepatocellular carcinoma using cancer-specific gene expression [J]. Cancer Res. 1995.55(22):5283-7.
[38]Ganslmayer M, Ocker M, Zopf S, et al. A quadruple therapy synergistically blocks proliferation and promotes apoptosis of hepatoma cells [J]. Oncol Rep. 2004.11(5):943-50.
[39]Li PY, Huang HJ, Zhang Q. [A study on the apoptosis of hepatoma cells synergistically induced by plasmid-mediated anti-angiogenesis and immunopotentiation therapy] [J]. Zhonghua Gan Zang Bing Za Zhi.2008.16(12): 909-12.
[40]Li PY, Lin JS, Feng ZH, et al. Combined gene therapy of endostatin and interleukin 12 with polyvinylpyrrolidone induces a potent antitumor effect on hepatoma [J]. World J Gastroenterol.2004.10(15):2195-200.
[41]Satoh H, Lamb PW, Dong JT, et al. Suppression of tumorigenicity of A549 lung adenocarcinoma cells by human chromosomes 3 and 11 introduced via microcell-mediated chromosome transfer [J]. Mol Carcinog.1993.7(3):157-64.
[42]Murakami Y, Nobukuni T, Tamura K, et al. Localization of tumor suppressor activity important in nonsmall cell lung carcinoma on chromosome 11q [J]. Proc Natl Acad Sci U S A.1998.95(14):8153-8.
[43]Murakami Y. Functional cloning of a tumor suppressor gene, TSLC1, in human non-small cell lung cancer [J]. Oncogene.2002.21(45):6936-48.
[44]Tsujiuchi T, Sugata E, Masaoka T, et al. Expression and DNA methylation patterns of Tslcl and Dal-1 genes in hepatocellular carcinomas induced by N-nitrosodiethylamine in rats [J]. Cancer Sci.2007.98(7):943-8.
[45]李文涛,白华龙,周闯,宋燕,冯留顺,张水军.原发性肝癌中抑癌基因TSLC1蛋白的表达及其临床意义[J].第四军医大学学报.2007.(23)：2141-2143.
[46]Iizuka M, Sugiyama Y, Shiraishi M, Jones C, Sekiya T. Allelic losses in human chromosome 11 in lung cancers [J]. Genes Chromosomes Cancer.1995.13(1):40-6.
[47]Gomyo H, Arai Y, Tanigami A, et al. A 2-Mb sequence-ready contig map and a novel immunoglobulin superfamily gene IGSF4 in the LOH region of chromosome Ilq23.2 [J]. Genomics.1999.62(2):139-46.
[48]Kuramochi M, Fukuhara H, Nobukuni T, et al. TSLC1 is a tumor-suppressor gene in human non-small-cell lung cancer [J]. Nat Genet.2001.27(4):427-30.
[49]Masuda M, Yageta M, Fukuhara H, et al. The tumor suppressor protein TSLC1 is involved in cell-cell adhesion [J]. Biol Chem.2002.277(34):31014-9.
[50]Ito A, Hagiyama M, Mimura T, et al. Expression of cell adhesion molecule 1 in malignant pleural mesothelioma as a cause of efficient adhesion and growth on mesothelium [J]. Lab Invest.2008.88(5):504-14.
[51]Koma Y, Furuno T, Hagiyama M, et al. Cell adhesion molecule 1 is a novel pancreatic-islet cell adhesion molecule that mediates nerve-islet cell interactions [J]. Gastroenterology.2008.134(5):1544-54.
[52]Fujita E, Soyama A, Momoi T. RA175, which is the mouse ortholog of TSLC1, a tumor suppressor gene in human lung cancer, is a cell adhesion molecule [J]. Exp Cell Res.2003.287(1):57-66.
[53]Watabe K, Ito A, Koma YI, Kitamura Y. IGSF4:a new intercellular adhesion molecule that is called by three names, TSLC1, SgIGSF and SynCAM, by virtue of its diverse function [J]. Histol Histopathol.2003.18(4):1321-9.
[54]Shingai T, Ikeda W, Kakunaga S, et al. Implications of nectin-like molecule-2/IGSF4/RA175/SgIGSF/TSLC1/SynCAMl in cell-cell adhesion and transmembrane protein localization in epithelial cells [J]. J Biol Chem.2003. 278(37):35421-7.
[55]Mandai K, Nakanishi H, Satoh A, et al. Afadin:A novel actin filament-binding protein with one PDZ domain localized at cadherin-based cell-to-cell adherens junction [J]. J Cell Biol.1997.139(2):517-28.
[56]Mandai K, Nakanishi H, Satoh A, et al. Ponsin/SH3P12:an 1-afadin-and vinculin-binding protein localized at cell-cell and cell-matrix adherens junctions [J]. J Cell Biol.1999.144(5):1001-17.
[57]Fukuhara H, Masuda M, Yageta M, et al. Association of a lung tumor suppressor TSLC1 with MPP3, a human homologue of Drosophila tumor suppressor Dlg [J]. Oncogene.2003.22(40):6160-5.
[58]Fukami T, Satoh H, Fujita E, et al. Identification of the Tslc1 gene, a mouse orthologue of the human tumor suppressor TSLC1 gene [J]. Gene.2002.295(1):7-12.
[59]Hoover KB, Bryant PJ. The genetics of the protein 4.1 family:organizers of the membrane and cytoskeleton [J]. Curr Opin Cell Biol.2000.12(2):229-34.
[60]Mangeat P, Roy C, Martin M. ERM proteins in cell adhesion and membrane dynamics [J]. Trends Cell Biol.1999.9(5):187-92.
[61]Tran YK, Bogler O, Gorse KM, Wieland I, Green MR, Newsham IF. A novel member of the NF2/ERM/4.1 superfamily with growth suppressing properties in lung cancer [J]. Cancer Res.1999.59(1):35-43.
[62]Galibert L, Diemer GS,Liu Z, et al. Nectin-like protein 2 defines a subset of T-cell zone dendritic cells and is a ligand for class-I-restricted T-cell-associated molecule [J]. J Biol Chem.2005.280(23):21955-64.
[63]Boles KS, Barchet W, Diacovo T, Cella M, Colonna M. The tumor suppressor TSLC1/NECL-2 triggers NK-cell and CD8+ T-cell responses through the cell-surface receptor CRTAM [J]. Blood.2005.106(3):779-86.
[64]Kakunaga S, Ikeda W, Itoh S, et al. Nectin-like molecule-1/TSLL1/SynCAM3:a neural tissue-specific immunoglobulin-like cell-cell adhesion molecule localizing at non-junctional contact sites of presynaptic nerve terminals, axons and glia cell processes [J]. J Cell Sci.2005.118(Pt 6):1267-77.
[65]Fukuhara H, Kuramochi M, Nobukuni T, et al. Isolation of the TSLL1 and TSLL2 genes, members of the tumor suppressor TSLC1 gene family encoding transmembrane proteins [J]. Oncogene.2001.20(38):5401-7.
[66]Fujita E, Kouroku Y, Ozeki S, et al. Oligo-astheno-teratozoospermia in mice lacking RA175/TSLC1/SynCAM/IGSF4A, a cell adhesion molecule in the immunoglobulin superfamily [J]. Mol Cell Biol.2006.26(2):718-26.
[67]Wakayama T, Koami H, Ariga H, et al. Expression and functional characterization of the adhesion molecule spermatogenic immunoglobulin superfamily in the mouse testis [J]. Biol Reprod.2003.68(5):1755-63.
[68]Ito A, Jippo T, Wakayama T, et al. SgIGSF:a new mast-cell adhesion molecule used for attachment to fibroblasts and transcriptionally regulated by MITF [J]. Blood.2003.101(7):2601-8.
[69]Biederer T. Bioinformatic characterization of the SynCAM family of immunoglobulin-like domain-containing adhesion molecules [J]. Genomics. 2006.87(1):139-50.
[70]Hirohashi S, Kanai Y. Cell adhesion system and human cancer morphogenesis [J]. Cancer Sci.2003.94(7):575-81.
[71]Mao X, Seidlitz E, Ghosh K, Murakami Y, Ghosh HP. The cytoplasmic domain is critical to the tumor suppressor activity of TSLC1 in non-small cell lung cancer [J]. Cancer Res.2003.63(22):7979-85.
[72]Yageta M, Kuramochi M, Masuda M, et al. Direct association of TSLC1 and DAL-1, two distinct tumor suppressor proteins in lung cancer [J]. Cancer Res. 2002.62(18):5129-33.
[73]Apoptosis [J]. Lancet.1972.2(7785):1011-2.
[74]da FRR, Kosiol C, Vinar T, Siepel A, Nielsen R. Positive selection on apoptosis related genes [J]. FEBS Lett.2010.584(3):469-76.
[75]Pop C, Salvesen GS. Human caspases:activation, specificity, and regulation [J]. J Biol Chem.2009.284(33):21777-81.
[76]Kurokawa M, Kornbluth S. Caspases and kinases in a death grip [J]. Cell.2009. 138(5):838-54.
[77]Mao X, Seidlitz E, Truant R, Hitt M, Ghosh HP. Re-expression of TSLC1 in a non-small-cell lung cancer cell line induces apoptosis and inhibits tumor growth [J]. Oncogene.2004.23(33):5632-42.
[78]Sussan TE, Pletcher MT, Murakami Y, Reeves RH. Tumor suppressor in lung cancer 1 (TSLC1) alters tumorigenic growth properties and gene expression [J]. Mol Cancer.2005.4:28.
[79]Jiang W, Newsham IF. The tumor suppressor DAL-1/4.1B and protein methylation cooperate in inducing apoptosis in MCF-7 breast cancer cells [J]. Mol Cancer.2006.5:4.
[80]Grimm M, Gasser M, Bueter M, et al. Evaluation of immunological escape mechanisms in a mouse model of colorectal liver metastases [J]. BMC Cancer. 2010.10:82.
[81]Palena C, Schlom J. Vaccines against Human Carcinomas:Strategies to Improve Antitumor Immune Responses [J]. Biomed Biotechnol.2010.2010:380697.
[82]Sasaki H, Nishikata I, Shiraga T, et al. Overexpression of a cell adhesion molecule, TSLC1, as a possible molecular marker for acute-type adult T-cell leukemia [J]. Blood.2005.105(3):1204-13.
[83]Dewan MZ, Takamatsu N, Hidaka T, et al. Critical role for TSLC1 expression in the growth and organ infiltration of adult T-cell leukemia cells in vivo [J]. J Virol. 2008.82(23):11958-63.
[84]Kitamura Y, Kurosawa G, Tanaka M, et al. Frequent overexpression of CADM1/IGSF4 in lung adenocarcinoma [J]. Biochem Biophys Res Commun. 2009.383(4):480-4.
[85]Fukuhara H, Kuramochi M, Fukami T, et al. Promoter methylation of TSLC1 and tumor suppression by its gene product in human prostate cancer [J]. Jpn J Cancer Res.2002.93(6):605-9.
[86]Jansen M, Fukushima N, Rosty C, et al. Aberrant methylation of the 5'CpG island of TSLC1 is common in pancreatic ductal adenocarcinoma and is first manifest in high-grade PanlNs [J]. Cancer Biol Ther.2002.1(3):293-6.
[87]Allinen M, Peri L, Kujala S, et al. Analysis of 11q21-24 loss of heterozygosity candidate target genes in breast cancer:indications of TSLC1 promoter hypermethylation [J]. Genes Chromosomes Cancer.2002.34(4):384-9.
[88]Steenbergen RD, Kramer D, Braakhuis BJ, et al. TSLC1 gene silencing in cervical cancer cell lines and cervical neoplasia [J]. J Natl Cancer Inst.2004. 96(4):294-305.
[89]Overmeer RM, Henken FE, Snijders PJ, et al. Association between dense CADM1 promoter methylation and reduced protein expression in high-grade CIN and cervical SCC [J]. J Pathol.2008.215(4):388-97.
[90]Fu L, Gao Z, Zhang X, et al. Frequent concomitant epigenetic silencing of the stress-responsive tumor suppressor gene CADM1, and its interacting partner DAL-1 in nasal NK/T-cell lymphoma [J]. Int J Cancer.2009.124(7):1572-8.
[91]Honda T, Tamura G, Waki T, et al. Hypermethylation of the TSLC1 gene promoter in primary gastric cancers and gastric cancer cell lines [J]. Jpn J Cancer Res.2002.93(8):857-60.
[92]Nowacki S, Skowron M, Oberthuer A, et al. Expression of the tumour suppressor gene CADM1 is associated with favourable outcome and inhibits cell survival in neuroblastoma [J]. Oncogene.2008.27(23):3329-38.
[93]Ando K, Ohira M, Ozaki T, et al. Expression of TSLC1, a candidate tumor suppressor gene mapped to chromosome 11q23, is downregulated in unfavorable neuroblastoma without promoter hypermethylation [J]. Int J Cancer.2008. 123(9):2087-94.
[94]Kikuchi S, Yamada D, Fukami T, et al. Hypermethylation of the TSLC1/IGSF4 promoter is associated with tobacco smoking and a poor prognosis in primary nonsmall cell lung carcinoma [J]. Cancer.2006.106(8):1751-8.
[95]Goto A, Niki T, Chi-Pin L, et al. Loss of TSLC1 expression in lung adenocarcinoma:relationships with histological subtypes, sex and prognostic significance [J]. Cancer Sci.2005.96(8):480-6.
[96]Fukami T, Fukuhara H, Kuramochi M, et al. Promoter methylation of the TSLC1 gene in advanced lung tumors and various cancer cell lines [J]. Int J Cancer. 2003.107(1):53-9.
[97]Uchino K, Ito A, Wakayama T, et al. Clinical implication and prognostic significance of the tumor suppressor TSLC1 gene detected in adenocarcinoma of the lung [J]. Cancer.2003.98(5):1002-7.
[98]Ito T, Shimada Y, Hashimoto Y, et al. Involvement of TSLC1 in progression of esophageal squamous cell carcinoma [J]. Cancer Res.2003.63(19):6320-6.
[99]Surace El, Lusis E, Murakami Y, Scheithauer BW, Perry A, Gutmann DH. Loss of tumor suppressor in lung cancer-1 (TSLC1) expression in meningioma correlates with increased malignancy grade and reduced patient survival [J]. J Neuropathol Exp Neurol.2004.63(10):1015-27.
[100]Shimizu K, Onishi M, Sugata E, Fujii H, Honoki K, Tsujiuchi T. Aberrant DNA methylation of the 5'upstream region of Tslc1 gene in hamster pancreatic tumors [J]. Biochem Biophys Res Commun.2007.353(2):522-6.
[101]Borinstein SC, Conerly M, Dzieciatkowski S, et al. Aberrant DNA methylation occurs in colon neoplasms arising in the azoxymethane colon cancer model [J]. Mol Carcinog.2010.49(1):94-103.
[102]袁林,陈安民,郭风劲,祝文涛,王江,廖晖.肺癌抑癌基因1对人前列腺癌T_3B细胞增殖的抑制效应[J].中国癌症杂志.2009.(01)：1-5.
[103]Heller G, Geradts J, Ziegler B, et al. Downregulation of TSLC1 and DAL-1 expression occurs frequently in breast cancer [J]. Breast Cancer Res Treat. 2007.103(3)：283-91.
[104]丁勇,李小荣,杨开焰,胡桂.乳腺癌组织中肺癌肿瘤抑制物1的表达及意义[J].实用预防医学.2009.(02)：355-357.
[105]Yang YX, Yang AH, Yang ZJ, Wang ZR, Xia XH. Involvement of tumor suppressor in lung cancer 1 gene expression in cervical carcinogenesis [J]. Int J Gynecol Cancer.2006.16(5):1868-72.
[106]Usami Y, Ito A, Ohnuma K, Fuku T, Komori T, Yokozaki H. Tumor suppressor in lung cancer-1 as a novel ameloblast adhesion molecule and its downregulation in ameloblastoma [J]. Pathol Int.2007.57(2):68-75.
[107]Hori RT. The minimal Tumor Suppressor in Lung Cancer-1 promoter is restrained by an inhibitory region [J]. Molecular Biology Reports.2009.
[108]Bhatia R, Dogra RC, Sharma PK. Construction of green fluorescent protein (GFP)-marked strains of Bradyrhizobium for ecological studies [J]. J Appl Microbiol.2002.93(5):835-9.
[109]Koczor CA, Snyder JW, Shokolenko IN, Dobson AW, Wilson GL, Ledoux SP. Targeting repair proteins to the mitochondria of mammalian cells through stable transfection, transient transfection, viral transduction, and TAT-mediated protein transduction [J]. Methods Mol Biol.2009.554: 233-49.
[110]Wright JF. Transient transfection methods for clinical adeno-associated viral vector production [J]. Hum Gene Ther.2009.20(7):698-706.
[111]Townsend-Nicholson A. Approaches to the stable transfection of G protein-coupled receptors [J]. Methods Mol Biol.1997.83:45-54.
[112]Douglas KL. Toward development of artificial viruses for gene therapy:a comparative evaluation of viral and non-viral transfection [J]. Biotechnol Prog.2008.24(4):871-83.
[113]Montier T, Benvegnu T, Jaffres PA, Yaouanc JJ, Lehn P. Progress in cationic lipid-mediated gene transfection:a series of bio-inspired lipids as an example [J]. Curr Gene Ther.2008.8(5):296-312.
[114]Susa T, Kato T, Kato Y. Reproducible transfection in the presence of carrier DNA using FuGENE6 and Lipofectamine2000 [J]. Mol Biol Rep.2008. 35(3):313-9.
[115]章静波.细胞生物学实用方法与技术[M].北京医科大学一中国协和医科 大学联合出版社.1995：581-597.
[116]Yamada D, Yoshida M, Williams YN, et al. Disruption of spermatogenic cell adhesion and male infertility in mice lacking TSLC1/IGSF4, an immunoglobulin superfamily cell adhesion molecule [J]. Mol Cell Biol.2006. 26(9):3610-24.
[117]Heller G, Fong KM, Girard L, et al. Expression and methylation pattern of TSLC1 cascade genes in lung carcinomas [J]. Oncogene.2006.25(6): 959-68.
[118]覃莉,张正茂,郝友华等.肺癌抑癌基因1对HepG2细胞生长的抑制效应[J].中华肝脏病杂志.2007.15(7)：509-512.