稳定转染靶向bcl-2的shRNA对胃癌SGC-7901细胞株5-FU/DDP敏感性的影响
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摘要
目的：探讨稳定转染靶向bcl-2的小发夹RNA对胃癌细胞株SGC-7901增殖、凋亡及化疗敏感性的影响。
方法：构建针对bcl-2的小发夹RNA质粒表达载体,转入SGC-7901细胞,筛选稳定转染的细胞克隆继续压力培养。不同浓度的5-Fu或DDP作用于稳定转染靶向Bcl-2 shRNA的胃癌SGC-7901细胞。RT-PCR法检测稳定转染后SGC-7901细胞bcl-2 mRNA的表达,MTT检测细胞增殖,流式细胞术检测细胞凋亡。
结果：稳定转染shRNA后,SGC-7901细胞的bcl-2 mRNA表达明显下降；细胞增殖能力及细胞凋亡率无明显变化。稳定转染组5-Fu的IC。。为14.36±1.63mg／L,未转染对照组为35.62±1.95 mg／L(p<0.05)；稳定转染组DDP的IC50为2.53±0.46mg／L,未转染对照组为6.89±0.52mg／L(p<0.05),流式细胞术显示稳定转染组凋亡率明显增高(p<0.05)。
结论：稳定转染shRNA能长效的抑制bcl-2 mRNA的表达,对细胞的增殖及凋亡无明显影响,能增加了5-Fu及DDP的化疗敏感性,为后续基因治疗研究提供实验依据。
Objective:To investigate the effect of stable expression shRNA targeted to bcl-2 in SGC-7901 cells.
Methods:A shRNA expressed vector that expresses the specific small hairpin RNA targeting bcl-2 mRNA was constructed and transfected into SGC-7901 cells. The stably expressing shRNA cells were selected by G418 and continuously cultured in half the antibiotic concentration, treated with 5-Fu or DDP. The bcl-2 mRNA was observed with RT-PCR, cellular proliferation capacity with MTT assay and the apoptotic state with flow cytometry.
Results:In SGC-7901 cells stably expressing shRNA, the expression of bcl-2 mRNA were decreased, but cellular proliferation capacity and apoptotic rate were normal. IC50 of 5-Fu was 14.36±1.63mg/L, IC50 of DDP was 2.53±0.46mg/L, apoptosis rate was increased.
Conclusion:The study demonstrated that shRNA has long-term effect to decrease the bcl-2 mRNA transcription and enhance the chemosensitivity, but cellular proliferation capacity and apoptotic rate were normal. which implies that regulate bcl-2 genes by RNAi may be a potential approach in cancer gene therapy.
方法：构建针对bcl-2的小发夹RNA质粒表达载体,转入SGC-7901细胞,筛选稳定转染的细胞克隆继续压力培养。不同浓度的5-Fu或DDP作用于稳定转染靶向Bcl-2 shRNA的胃癌SGC-7901细胞。RT-PCR法检测稳定转染后SGC-7901细胞bcl-2 mRNA的表达,MTT检测细胞增殖,流式细胞术检测细胞凋亡。
结果：稳定转染shRNA后,SGC-7901细胞的bcl-2 mRNA表达明显下降；细胞增殖能力及细胞凋亡率无明显变化。稳定转染组5-Fu的IC。。为14.36±1.63mg／L,未转染对照组为35.62±1.95 mg／L(p<0.05)；稳定转染组DDP的IC50为2.53±0.46mg／L,未转染对照组为6.89±0.52mg／L(p<0.05),流式细胞术显示稳定转染组凋亡率明显增高(p<0.05)。
结论：稳定转染shRNA能长效的抑制bcl-2 mRNA的表达,对细胞的增殖及凋亡无明显影响,能增加了5-Fu及DDP的化疗敏感性,为后续基因治疗研究提供实验依据。
Objective:To investigate the effect of stable expression shRNA targeted to bcl-2 in SGC-7901 cells.
Methods:A shRNA expressed vector that expresses the specific small hairpin RNA targeting bcl-2 mRNA was constructed and transfected into SGC-7901 cells. The stably expressing shRNA cells were selected by G418 and continuously cultured in half the antibiotic concentration, treated with 5-Fu or DDP. The bcl-2 mRNA was observed with RT-PCR, cellular proliferation capacity with MTT assay and the apoptotic state with flow cytometry.
Results:In SGC-7901 cells stably expressing shRNA, the expression of bcl-2 mRNA were decreased, but cellular proliferation capacity and apoptotic rate were normal. IC50 of 5-Fu was 14.36±1.63mg/L, IC50 of DDP was 2.53±0.46mg/L, apoptosis rate was increased.
Conclusion:The study demonstrated that shRNA has long-term effect to decrease the bcl-2 mRNA transcription and enhance the chemosensitivity, but cellular proliferation capacity and apoptotic rate were normal. which implies that regulate bcl-2 genes by RNAi may be a potential approach in cancer gene therapy.
引文
[1]Antonsson B and Martinou JC. The Bcl-2 protein family[J]. Exp Cell Res,2000,256: 50.
[2]Jian Y, Lin ZB. Apoptosis in human cancer cells[J]. Current Opinion in Oncology,2004; 16:19-24.
[3]Adams JM, Cory S. The bcl-2 protein family:arbiers of cell survival[J]. Science, 2004,281:1322.
[4]FIRE A, XU S, MONTGOMERY MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenothabditls elegans[J]. Nature,1998,391:808-811.
[5]SuzukiC, DaigoY, KikuchiTetal. Identification of Cox17 as a therapeutic target for non-small cell lung cancer[J]. CancerRes,2003; 63(21):7038-41.
[6]Hong J H, Lee E, Hong J, et al. Antisense Bcl2 oligonucleotide in cisplatin-resistant bladder cancer cell lines. BJ U Int,2002,90:113-117.
[7]Cioca DP, Aoki Y, Kiyosawa K. RNA interference is a functional pathway with therapeutic potential in human myeloid leukemia cell lines [M]. Cancer Gene Ther,2003,10(2):125-133.
[8]Van Dedonk NW, Kamphui MM, Van Diikm, et al. Chemosensitization of myeloma plasma cells by an antisense-mediated downregulation of Bcl-2 protein[J]. Leukemia,2003; 17(1): 211-219.
[9]Marcucci G, Byrd JC, Dai G, et al. Phase 1 and pharmacodynamic studies of G3139, a Bcl-2 antisense oligonucleotide, in combination with chemotherapy in refractory or relapsed acute leukemia[J]. Blood,2003; 101(2):425-432.
[10]Waters JS, Webb A, Cunningham D, et al. Phase I clinical and pharmacokinetic study of Bcl-2 antisense oligonucleotide therapy in patients with non-Hodgkin's lymphoma[J]. J Clin Oncol.2000; 18(9):1812-1823.
[11]Holen T, Amarzguioui M, Wiiger MT, et al. Positional effects of short interfering RNAs targeting the human coagulation trigger Tissue Factor. Nuclei Acids Res.2002; 30(8): 1757-1766.
[12]YIN JQ, GAO GS, SHAO RG, et al. siRNA agents inhibit oncogene expression and attenuate human tumor cell growth [J]. Exp Ther Oncol,2003,3(4):194-204.
[13]BRUM MELKAMP TR, BERNARDS R, AGAMI R. Stable suppression of tumorigenicity by virus-mediated RNA interference[J]. Cancer Cell,2002,2(3):234-247.
[14]Rubinson DA, Dillon CP, Kwiatkowski AV, et al. A lentivirus—based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference [J]. Nat Genet,2003,33(3):401—406.
[15]王明玉,宋丽华,宋现让,等.人乳腺癌MDR1基因稳定沉默细胞的筛选及其生物学特性[J].基础医学与临床,2008,28(2)：138-143.
[16]裘莹,宋伯根,沈丽贤.Bcl-2蛋白与胃癌相关性的研究[J].同济大学学报,2001,22(1)：16-17.
[17]吴炅,沈镇宙.乳腺癌中Bcl-2基因族研究现状[J].中国癌症杂志,1998；8(3)：217-220.
218]Kim R, Emi M, Tanabe K, et al. Preclinical evaluation of antisense bcl-2 as a chemosensitizer for patients with gastric carcinoma. Cancer,2004;101:2177-2186.
[19]Kim R, Emi M,Matsuura K, et al. Therapeutic potential of antisense (AS) Bcl-2 as a chemosensitizer for patients with gastriand breast carcinoma. Gan To Kagaku Ryoho,2005; 32:1540-1545.
[20]Wacheck V, Heere-Ress E, Halaschek-Wiener J, et al. Bcl-2 antisense oligonucleotides chemosensitize human gastric cancer in SCID mouse xenotransplantation model. J Mol Med, 2001; 79:587-593.
[21]Holle L, Hicks L, Songw, et al. Bcl-2 targeting siRNA expressed by a T7 vector system inhibits human tumor cell growth in vitro[J]. Int J Oncol,2004,24:615-621.
[22]Uchida H, Tanaka T, Sasaki K, et al. Adenovirus-mediated transfer of siRNA against survivin induced apoptosis and attenuated tumor cell growth in vitro and in vivo[J]. Mol Ther,2004.10:162-171.
[23]BRUMMELKAMP TR, BERNARDS R, AGAMI R. A system for stable expression of short interfering RNAs in mammalian cells[J]. Science,2002,296:550-553.
[24]许德晖,黄辰,刘利英,等.高效siRNA设计的研究进展.遗传,2006,28(11)：1457-1461.
1 Hannon GJ. RNA interference. Nature,2002,418(6894):244-251.
2 Liu N, Bi F, Pan Y, el al. Reversal of the malignant phenotype of gastric cancer cells by inhibition of RhoA expression and activity. Clin Cancer Res,2004,10:6239-6247.
3刘娜,毕锋,潘阳林,等.RhoC在胃癌细胞中的表达及其小干扰RNA表达载体的构建与鉴定.细胞与分子免疫学杂志,2004,20：148-151.
4 Jang YJ, Kim YS, Kim WH. Oncogenic effect of Polo-like kinase 1 expression in human gastric carcinomas. Int Oncol,2006,29(3):589-594.
5Jia SC, Su CQ, Zhang WB et al. Effects of survivin gene siRNA on the growth of gastric cancer cell line [J].Chin J Gen Sur,2007,22(12):925-927.
6 Fu GF, Lin XH, Han QW, et al. RNA interference remarkably suppresses bcl-2 gene expression in cancer cells in vitro and in vivo[J]. Cancer Biol Ther,2005,4:822-829.
7 Nieth C, Priebsch A, Stege A, et al. Modulation of the classical multidrug resistance (MDR) phenotype by RNA interference(RNAi). FEBS Lett,2003,545(3):144-150.
8 Hong L, Ning X, Shi Y, et al. Reversal of multidrug resistance of gastric cancer cells by down-regulation of ZNRD1 with ZNRD1 siRNA.Br J Biomed Sci,2004,61(4):206-210.
9 Yang YX, Xiao ZQ, Chen ZC, et al. Proteme analysis of multidrug resistance in vincristine-resistant human gastric cancer cell lineSGC7901/VCR[J]. Proteomics,2006,6(6): 2009-2021.
10 He QC, Zhang GY, Cao WJ. Correlation of Sorcin Overexpression to Multidrug Resistance of Human Gastric Cancer Cell Line SGC7901[J].Chin J Cancer,2008,27(4):337-342.
11 Ito R, Nakayama H, Yoshida K, et al. Expression of Cbl linking with the epidermal growth factor receptor system is assoeiated with tumor progression and poor prognosis of human gastric carcinoma. Virchows Arch,2004,444:324-331.
12 Saukkonen K, Rintahaka J, Sivula A, et al. Cyclooxygenase-2 and gastric carcinogenesis. APMIS,2003,111(10):915-925.
13李玉明,詹文华,蔡世荣,等.靶向环氧合酶-2的RNA干扰诱导胃癌SGC-7901细胞凋亡.中华实验外科杂志,2006,23：662-664.
14 Jinawath N, Furukawa Y, Nakamura Y. Identification of NOL1 8, a nucleolar protein containing an RNA recoguition motif(RRM), which was overexpressed in diffusetype gastric cancer[J]. Cancer Sci,2004,95(5):430-435.
15 Choudhury A, Charo J, Parapuram SK, et al. Small interfering RNA (siRNA) inhibits the expression of the Her2/neu gene, upregulates HLA class I and induces apoptosis of Her2 /neu positive tumor cell lines. Int J Cancer,2004,108:71-77.
16 Jiang Y, Wang L, Gong W, et al.A high expression level of Insulin-like growth factor I receptor is associated with increased expression of transcription factor Spl and regional lymph node metastasis of human gastric cancer. Clin Exp Metastasis,2004,21(8):755-764.
17 Yan C, Zhu ZG, Yu YY, et al. Expression of vascular endothelial growth factor C and chemokine receptor CCR7 in gastric carcinoma and their values in predicting lymph node metastasis. World J Gastroenterol,2004,10:783-790.
18 Filleur S, Courtin A, Ait Si Ali S, et al. SiRNA-mediated inhibition of vascular endothelial growth factor severely limits tumor resistance to antiangiogenic thrombospondin-1 and slows tumor vascularization and growth. Cancer Res,2003,63:3919-3922.
19徐文华,葛银林,徐宏伟,等.VEGF基因表达抑制对胃腺癌细胞SGC-7901增殖的影响.世界华人消化杂志,2006,14(7)：655-659.
20 Meng F, Ding J, Liu N, et al. Inhibition of gastric cancer angiogenesis by vector-based RNA interference for Raf-1. Cancer Biol Ther,2005,4(1):113-117.
21 Xue Y, Bi F, Zhang X, et al. Inhibition of endothelial cell proliferation by targeting Racl GTPase with small interference RNA in tumor cells. Biochem Biophys Res Commun.2004, 320:1309-1315.
22吴涛,徐惠棉,吴晓华,等.应用载体介导的RNAi技术抑制人胃癌和腹膜间皮细胞中TGF-β1的表达[J].世界华人消化杂志,2005,13(21)：2530-2534.
23 Tsuchiya T, Okaji Y, Tsuno NH, el al. Targeting Idl and Id3 inhibits peritoneal metastasis of gastric cancer[J].Cancer Sci,2005,96(11):784-790.
24 Zheng ZH, Sun XJ, Zhou HT, et al. Analysis of metastasis suppressing function of E-cadherin in gastric cancer cells by RNAi[J]. World J Gastroenteral,2005,Ⅱ(13):2000-2003.
25 SHIN J, KIM J, RYU B, et al. Caveolin-1 is associated with VCAM-1 dependent adhesion of gastric cancer cells to endothdial cells[J].Cel Physiol Biochem,2006,17(5-6):211.
26李莹,药立波,刘新平,等.RNAi引起的Paxillin和p130Cas下调抑制胃癌细胞失巢性生长[J].中国生物化学与分子生物学报,2005,21(3)：408.
27 DENG H, GUO RF, L1 WM, et al. Matrix metalloproteinase 11 depletion inhibits cell proliferation in gastric cancer cells[J]. Biochem biophys Res Commun,2005,325(2):274.
28 Collis SJ, Swartz MJ, Nelson WG, et al. Enhanced radiation and chemotherapy-mediated cell killing of human cancer cells by small inhibitory RNA silencing of DNA repair factors [J]. Cancer Res,2003,63(7):1550-1554.
29 Peng Y, Zhang Q, Nagasawa H, et al. Silencing expression of the catalytic subunit of DNA-dependent protein kinase by small interfering RNA sensitizes human cells for radiation-induced chromosome damage, cell killing, and mutation[J]. Cancer Res, 2002,62(22):6400-6404.
30 The Thomson Corporation, Thomson Scientifics review of phase changes in the pharmaceutical pipeline[R]. The Ones to Watch,2007:3-4.
31 Song HF, Zhou J, Pan K, et al. Silence SOCS1 by siRNA, Stimulated by OK-432 and Pulsed with Lysate of HepG2 Cells[J]. Chinese J Cancer,2008,27(7):685-691.
32 Weng DH, Wang SX, Ma D. Application of RNAi Library to Oncology Investigation [J]. Chinese J Cancer,2008,27 (11):1229-1232.
[2]Jian Y, Lin ZB. Apoptosis in human cancer cells[J]. Current Opinion in Oncology,2004; 16:19-24.
[3]Adams JM, Cory S. The bcl-2 protein family:arbiers of cell survival[J]. Science, 2004,281:1322.
[4]FIRE A, XU S, MONTGOMERY MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenothabditls elegans[J]. Nature,1998,391:808-811.
[5]SuzukiC, DaigoY, KikuchiTetal. Identification of Cox17 as a therapeutic target for non-small cell lung cancer[J]. CancerRes,2003; 63(21):7038-41.
[6]Hong J H, Lee E, Hong J, et al. Antisense Bcl2 oligonucleotide in cisplatin-resistant bladder cancer cell lines. BJ U Int,2002,90:113-117.
[7]Cioca DP, Aoki Y, Kiyosawa K. RNA interference is a functional pathway with therapeutic potential in human myeloid leukemia cell lines [M]. Cancer Gene Ther,2003,10(2):125-133.
[8]Van Dedonk NW, Kamphui MM, Van Diikm, et al. Chemosensitization of myeloma plasma cells by an antisense-mediated downregulation of Bcl-2 protein[J]. Leukemia,2003; 17(1): 211-219.
[9]Marcucci G, Byrd JC, Dai G, et al. Phase 1 and pharmacodynamic studies of G3139, a Bcl-2 antisense oligonucleotide, in combination with chemotherapy in refractory or relapsed acute leukemia[J]. Blood,2003; 101(2):425-432.
[10]Waters JS, Webb A, Cunningham D, et al. Phase I clinical and pharmacokinetic study of Bcl-2 antisense oligonucleotide therapy in patients with non-Hodgkin's lymphoma[J]. J Clin Oncol.2000; 18(9):1812-1823.
[11]Holen T, Amarzguioui M, Wiiger MT, et al. Positional effects of short interfering RNAs targeting the human coagulation trigger Tissue Factor. Nuclei Acids Res.2002; 30(8): 1757-1766.
[12]YIN JQ, GAO GS, SHAO RG, et al. siRNA agents inhibit oncogene expression and attenuate human tumor cell growth [J]. Exp Ther Oncol,2003,3(4):194-204.
[13]BRUM MELKAMP TR, BERNARDS R, AGAMI R. Stable suppression of tumorigenicity by virus-mediated RNA interference[J]. Cancer Cell,2002,2(3):234-247.
[14]Rubinson DA, Dillon CP, Kwiatkowski AV, et al. A lentivirus—based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference [J]. Nat Genet,2003,33(3):401—406.
[15]王明玉,宋丽华,宋现让,等.人乳腺癌MDR1基因稳定沉默细胞的筛选及其生物学特性[J].基础医学与临床,2008,28(2)：138-143.
[16]裘莹,宋伯根,沈丽贤.Bcl-2蛋白与胃癌相关性的研究[J].同济大学学报,2001,22(1)：16-17.
[17]吴炅,沈镇宙.乳腺癌中Bcl-2基因族研究现状[J].中国癌症杂志,1998；8(3)：217-220.
218]Kim R, Emi M, Tanabe K, et al. Preclinical evaluation of antisense bcl-2 as a chemosensitizer for patients with gastric carcinoma. Cancer,2004;101:2177-2186.
[19]Kim R, Emi M,Matsuura K, et al. Therapeutic potential of antisense (AS) Bcl-2 as a chemosensitizer for patients with gastriand breast carcinoma. Gan To Kagaku Ryoho,2005; 32:1540-1545.
[20]Wacheck V, Heere-Ress E, Halaschek-Wiener J, et al. Bcl-2 antisense oligonucleotides chemosensitize human gastric cancer in SCID mouse xenotransplantation model. J Mol Med, 2001; 79:587-593.
[21]Holle L, Hicks L, Songw, et al. Bcl-2 targeting siRNA expressed by a T7 vector system inhibits human tumor cell growth in vitro[J]. Int J Oncol,2004,24:615-621.
[22]Uchida H, Tanaka T, Sasaki K, et al. Adenovirus-mediated transfer of siRNA against survivin induced apoptosis and attenuated tumor cell growth in vitro and in vivo[J]. Mol Ther,2004.10:162-171.
[23]BRUMMELKAMP TR, BERNARDS R, AGAMI R. A system for stable expression of short interfering RNAs in mammalian cells[J]. Science,2002,296:550-553.
[24]许德晖,黄辰,刘利英,等.高效siRNA设计的研究进展.遗传,2006,28(11)：1457-1461.
1 Hannon GJ. RNA interference. Nature,2002,418(6894):244-251.
2 Liu N, Bi F, Pan Y, el al. Reversal of the malignant phenotype of gastric cancer cells by inhibition of RhoA expression and activity. Clin Cancer Res,2004,10:6239-6247.
3刘娜,毕锋,潘阳林,等.RhoC在胃癌细胞中的表达及其小干扰RNA表达载体的构建与鉴定.细胞与分子免疫学杂志,2004,20：148-151.
4 Jang YJ, Kim YS, Kim WH. Oncogenic effect of Polo-like kinase 1 expression in human gastric carcinomas. Int Oncol,2006,29(3):589-594.
5Jia SC, Su CQ, Zhang WB et al. Effects of survivin gene siRNA on the growth of gastric cancer cell line [J].Chin J Gen Sur,2007,22(12):925-927.
6 Fu GF, Lin XH, Han QW, et al. RNA interference remarkably suppresses bcl-2 gene expression in cancer cells in vitro and in vivo[J]. Cancer Biol Ther,2005,4:822-829.
7 Nieth C, Priebsch A, Stege A, et al. Modulation of the classical multidrug resistance (MDR) phenotype by RNA interference(RNAi). FEBS Lett,2003,545(3):144-150.
8 Hong L, Ning X, Shi Y, et al. Reversal of multidrug resistance of gastric cancer cells by down-regulation of ZNRD1 with ZNRD1 siRNA.Br J Biomed Sci,2004,61(4):206-210.
9 Yang YX, Xiao ZQ, Chen ZC, et al. Proteme analysis of multidrug resistance in vincristine-resistant human gastric cancer cell lineSGC7901/VCR[J]. Proteomics,2006,6(6): 2009-2021.
10 He QC, Zhang GY, Cao WJ. Correlation of Sorcin Overexpression to Multidrug Resistance of Human Gastric Cancer Cell Line SGC7901[J].Chin J Cancer,2008,27(4):337-342.
11 Ito R, Nakayama H, Yoshida K, et al. Expression of Cbl linking with the epidermal growth factor receptor system is assoeiated with tumor progression and poor prognosis of human gastric carcinoma. Virchows Arch,2004,444:324-331.
12 Saukkonen K, Rintahaka J, Sivula A, et al. Cyclooxygenase-2 and gastric carcinogenesis. APMIS,2003,111(10):915-925.
13李玉明,詹文华,蔡世荣,等.靶向环氧合酶-2的RNA干扰诱导胃癌SGC-7901细胞凋亡.中华实验外科杂志,2006,23：662-664.
14 Jinawath N, Furukawa Y, Nakamura Y. Identification of NOL1 8, a nucleolar protein containing an RNA recoguition motif(RRM), which was overexpressed in diffusetype gastric cancer[J]. Cancer Sci,2004,95(5):430-435.
15 Choudhury A, Charo J, Parapuram SK, et al. Small interfering RNA (siRNA) inhibits the expression of the Her2/neu gene, upregulates HLA class I and induces apoptosis of Her2 /neu positive tumor cell lines. Int J Cancer,2004,108:71-77.
16 Jiang Y, Wang L, Gong W, et al.A high expression level of Insulin-like growth factor I receptor is associated with increased expression of transcription factor Spl and regional lymph node metastasis of human gastric cancer. Clin Exp Metastasis,2004,21(8):755-764.
17 Yan C, Zhu ZG, Yu YY, et al. Expression of vascular endothelial growth factor C and chemokine receptor CCR7 in gastric carcinoma and their values in predicting lymph node metastasis. World J Gastroenterol,2004,10:783-790.
18 Filleur S, Courtin A, Ait Si Ali S, et al. SiRNA-mediated inhibition of vascular endothelial growth factor severely limits tumor resistance to antiangiogenic thrombospondin-1 and slows tumor vascularization and growth. Cancer Res,2003,63:3919-3922.
19徐文华,葛银林,徐宏伟,等.VEGF基因表达抑制对胃腺癌细胞SGC-7901增殖的影响.世界华人消化杂志,2006,14(7)：655-659.
20 Meng F, Ding J, Liu N, et al. Inhibition of gastric cancer angiogenesis by vector-based RNA interference for Raf-1. Cancer Biol Ther,2005,4(1):113-117.
21 Xue Y, Bi F, Zhang X, et al. Inhibition of endothelial cell proliferation by targeting Racl GTPase with small interference RNA in tumor cells. Biochem Biophys Res Commun.2004, 320:1309-1315.
22吴涛,徐惠棉,吴晓华,等.应用载体介导的RNAi技术抑制人胃癌和腹膜间皮细胞中TGF-β1的表达[J].世界华人消化杂志,2005,13(21)：2530-2534.
23 Tsuchiya T, Okaji Y, Tsuno NH, el al. Targeting Idl and Id3 inhibits peritoneal metastasis of gastric cancer[J].Cancer Sci,2005,96(11):784-790.
24 Zheng ZH, Sun XJ, Zhou HT, et al. Analysis of metastasis suppressing function of E-cadherin in gastric cancer cells by RNAi[J]. World J Gastroenteral,2005,Ⅱ(13):2000-2003.
25 SHIN J, KIM J, RYU B, et al. Caveolin-1 is associated with VCAM-1 dependent adhesion of gastric cancer cells to endothdial cells[J].Cel Physiol Biochem,2006,17(5-6):211.
26李莹,药立波,刘新平,等.RNAi引起的Paxillin和p130Cas下调抑制胃癌细胞失巢性生长[J].中国生物化学与分子生物学报,2005,21(3)：408.
27 DENG H, GUO RF, L1 WM, et al. Matrix metalloproteinase 11 depletion inhibits cell proliferation in gastric cancer cells[J]. Biochem biophys Res Commun,2005,325(2):274.
28 Collis SJ, Swartz MJ, Nelson WG, et al. Enhanced radiation and chemotherapy-mediated cell killing of human cancer cells by small inhibitory RNA silencing of DNA repair factors [J]. Cancer Res,2003,63(7):1550-1554.
29 Peng Y, Zhang Q, Nagasawa H, et al. Silencing expression of the catalytic subunit of DNA-dependent protein kinase by small interfering RNA sensitizes human cells for radiation-induced chromosome damage, cell killing, and mutation[J]. Cancer Res, 2002,62(22):6400-6404.
30 The Thomson Corporation, Thomson Scientifics review of phase changes in the pharmaceutical pipeline[R]. The Ones to Watch,2007:3-4.
31 Song HF, Zhou J, Pan K, et al. Silence SOCS1 by siRNA, Stimulated by OK-432 and Pulsed with Lysate of HepG2 Cells[J]. Chinese J Cancer,2008,27(7):685-691.
32 Weng DH, Wang SX, Ma D. Application of RNAi Library to Oncology Investigation [J]. Chinese J Cancer,2008,27 (11):1229-1232.