原发性肝癌组织中肝癌干细胞的检测及临床意义
摘要
肿瘤干细胞是最近几年提出的关于肿瘤发生、发展机制的新理论,该理论认为,导致肿瘤发生和维持肿瘤生长的是一小群叫做“肿瘤干细胞”的细胞,这些细胞在肿瘤组织中数量极少,具有自我更新、分化及抗治疗能力等干细胞特性,影响着肿瘤的进展、复发和转移,只有消灭这些细胞才能根治肿瘤。CD133是一种较新的肝癌干细胞的检查指标;血清甲胎蛋白(AFP)检测是当前诊断肝癌常用而又重要的方法,于肝癌干细胞中亦有表达。本文主要研究了肝癌干细胞标记物CD133及AFP在肝细胞肝癌中的表达及其意义并探讨其于预后的相关性。
实验目的：CD133及AFP在肝细胞肝癌中的表达及与肿瘤的生物学特性和术后生存期的影响。
实验方法：收集山东省立医院肝胆胰脾外科2004年4月至2006年5因肝细胞肝癌而行肝癌切除术的肝癌组织,共88例,用免疫组织化学SABC三步法检测肝癌组织中的CD133及AFP的表达,并研究CD133阳性表达及AFP阳性表达与临床病理联系(运用X2检验,P<0.05具有统计学意义),并对CD133阳性表达及AFP阳性表达对术后1年生存率与无瘤生存率及5年生存率与无瘤生存率的影响进行比较分析(运用Kaplan-Meier生存曲线)。
结果：
1.88例肝癌组织中CD133的阳性表达数为45例,阳性表达率为51.13%。
2.88例肝癌组织中AFP的阳性表达数为57例,阳性表达率为64.77%。
3.CD133及AFP同时阳性表达数为37例,P<0.05,CD133阳性表达与AFP阳性表达呈正相关。
4.CD133阳性表达及AFP的阳性表达均与患者的年龄、性别、饮酒史,血清HbsAg、肝硬化及肿瘤的大小无关(P>0.05),与肿瘤数目、血清AFP水平、分化程度及肿瘤分期有关(P<0.05)。
5. Kaplan-Meier生存曲线法发现CD133阳性表达及AFP阳性表达较CD133阴性表达及AFP阴性表达具有更短的1年和5年无瘤生存时间(P<0.05)。CD133阳性表达及AFP阳性表达较CD133阴性表达及AFP阴性表达具有更短的5年生存时间(P<0.05),但是1年生存时间无差异(P>0.05)。
结论：CD133及AFP在肝细胞肝癌中有表达,两者具有相关性,具有肿瘤干细胞特性,其表达与肿瘤的恶性程度及术后生存率有关,可以作为肝细胞肝癌愈后的指标。
Cancer stem cells is a new theory which appears in recent years about the occurrence and development of tumor. According to this theory, the factor that cause the occurrence and keep the development of tumor is a group of cells called "Cancer stem cells". These cells are very few in the tumor tissue, but they have the features of liver cells such as self-renewal, differentiation and anti-treatment, and they can affect the progression, recurrence and metastasis of the tumor. Only by eliminating these cells can we cure cancer. CD133 is a new indicators of liver cancer stem cells; Serum alpha-fetoprotein (AFP) test is one common and important diagnostic method of liver cancer so far, it is also expressed in liver cancer stem cells. This article studied the expression and significance of CD133 and AFP in hepatocellular carcinoma and investigate their relationship with prognosis.
OBJECTIVE:The expression of CD133 and AFP in hepatocellular carcinoma and their effect on the survival curve after surgery.
METHODS:Collection of 88 liver tumor tissues that are removed through the tumor resection for hepatocellular carcinoma in the Pancreatic and splenic department of Shandong Provincial Hospital from April 2003 to May 2005. Select the three-step method of SABC of immunohistochemistry to detect the expression of CD133 and AFP in liver tumor tissues, to investigate the relationship between the positive expression of CD133 and AFP and clinical pathology (select X2 test, P<0.05 has statistical significance), and comparatively analyze the effect of the positive expression of CD133 and AFP to the one-year survival rate and disease free survival rate and five-year survival rate and disease free survival rate after surgery (select the Kaplan-Meier survival curve).
RESULTS:
1.45 patients (51.13%) of 88 cases had the positive expression of CD133.
2.57 patients (64.77%) of 88 cases had the positive expression of AFP.
3.37 cases had the positive expression of both CD133 and AFP, P<0.05, the positive expression of CD133 and AFP had positive correlation.
4. Positive expression of CD133 and AFP was unrelated to patient's age, gender, alcohol consumption, serum HbsAg, liver cirrhosis and tumor size(P>0.05), but related to the number of tumors, serum AFP level, differentiation and tumor stage(P<0.05).
5. Kaplan-Meier survival curve showed that patients with positive expression of CD133 and AFP had shorter one-year and five-year free survival time than that with negative expression(P<0.05), and patients with positive expression of CD133 and AFP had shorter five-year survival time than that with negative expression(P<0.05), but no difference in one-year survival time(P>0.05).
CONCLUSIONS:
The proteins of CD133 and AFP were proved to be expressed in hepatocellular carcinoma. The two relevant have the properties of cancer stem cell, and their expression are related to the malignancy of tumor and postoperative survival rate. Therefore, they can be used as the indicators of prognosis of hepatocellular carcinoma.
实验目的：CD133及AFP在肝细胞肝癌中的表达及与肿瘤的生物学特性和术后生存期的影响。
实验方法：收集山东省立医院肝胆胰脾外科2004年4月至2006年5因肝细胞肝癌而行肝癌切除术的肝癌组织,共88例,用免疫组织化学SABC三步法检测肝癌组织中的CD133及AFP的表达,并研究CD133阳性表达及AFP阳性表达与临床病理联系(运用X2检验,P<0.05具有统计学意义),并对CD133阳性表达及AFP阳性表达对术后1年生存率与无瘤生存率及5年生存率与无瘤生存率的影响进行比较分析(运用Kaplan-Meier生存曲线)。
结果：
1.88例肝癌组织中CD133的阳性表达数为45例,阳性表达率为51.13%。
2.88例肝癌组织中AFP的阳性表达数为57例,阳性表达率为64.77%。
3.CD133及AFP同时阳性表达数为37例,P<0.05,CD133阳性表达与AFP阳性表达呈正相关。
4.CD133阳性表达及AFP的阳性表达均与患者的年龄、性别、饮酒史,血清HbsAg、肝硬化及肿瘤的大小无关(P>0.05),与肿瘤数目、血清AFP水平、分化程度及肿瘤分期有关(P<0.05)。
5. Kaplan-Meier生存曲线法发现CD133阳性表达及AFP阳性表达较CD133阴性表达及AFP阴性表达具有更短的1年和5年无瘤生存时间(P<0.05)。CD133阳性表达及AFP阳性表达较CD133阴性表达及AFP阴性表达具有更短的5年生存时间(P<0.05),但是1年生存时间无差异(P>0.05)。
结论：CD133及AFP在肝细胞肝癌中有表达,两者具有相关性,具有肿瘤干细胞特性,其表达与肿瘤的恶性程度及术后生存率有关,可以作为肝细胞肝癌愈后的指标。
Cancer stem cells is a new theory which appears in recent years about the occurrence and development of tumor. According to this theory, the factor that cause the occurrence and keep the development of tumor is a group of cells called "Cancer stem cells". These cells are very few in the tumor tissue, but they have the features of liver cells such as self-renewal, differentiation and anti-treatment, and they can affect the progression, recurrence and metastasis of the tumor. Only by eliminating these cells can we cure cancer. CD133 is a new indicators of liver cancer stem cells; Serum alpha-fetoprotein (AFP) test is one common and important diagnostic method of liver cancer so far, it is also expressed in liver cancer stem cells. This article studied the expression and significance of CD133 and AFP in hepatocellular carcinoma and investigate their relationship with prognosis.
OBJECTIVE:The expression of CD133 and AFP in hepatocellular carcinoma and their effect on the survival curve after surgery.
METHODS:Collection of 88 liver tumor tissues that are removed through the tumor resection for hepatocellular carcinoma in the Pancreatic and splenic department of Shandong Provincial Hospital from April 2003 to May 2005. Select the three-step method of SABC of immunohistochemistry to detect the expression of CD133 and AFP in liver tumor tissues, to investigate the relationship between the positive expression of CD133 and AFP and clinical pathology (select X2 test, P<0.05 has statistical significance), and comparatively analyze the effect of the positive expression of CD133 and AFP to the one-year survival rate and disease free survival rate and five-year survival rate and disease free survival rate after surgery (select the Kaplan-Meier survival curve).
RESULTS:
1.45 patients (51.13%) of 88 cases had the positive expression of CD133.
2.57 patients (64.77%) of 88 cases had the positive expression of AFP.
3.37 cases had the positive expression of both CD133 and AFP, P<0.05, the positive expression of CD133 and AFP had positive correlation.
4. Positive expression of CD133 and AFP was unrelated to patient's age, gender, alcohol consumption, serum HbsAg, liver cirrhosis and tumor size(P>0.05), but related to the number of tumors, serum AFP level, differentiation and tumor stage(P<0.05).
5. Kaplan-Meier survival curve showed that patients with positive expression of CD133 and AFP had shorter one-year and five-year free survival time than that with negative expression(P<0.05), and patients with positive expression of CD133 and AFP had shorter five-year survival time than that with negative expression(P<0.05), but no difference in one-year survival time(P>0.05).
CONCLUSIONS:
The proteins of CD133 and AFP were proved to be expressed in hepatocellular carcinoma. The two relevant have the properties of cancer stem cell, and their expression are related to the malignancy of tumor and postoperative survival rate. Therefore, they can be used as the indicators of prognosis of hepatocellular carcinoma.
引文
1 Parkin DM, Bray F, Ferlay J, Pisani P:Estimating the world cancer burden: Globocan 2000. Int J Cancer 2001,94:153-6.
2.Perz JF, Armstrong GL, Farrington LA, Hutin YJ, Bell BP:The contributions of hepatitis B virus and hepatitis C virus infections to cirrhosis and primary liver cancer worldwide. J Hepatol 2006,45:529-38.
3.Tanaka K, Hirohata T, Takeshita S, Hirohata I, Koga S, Sugimachi K,Kanematsu T, Ohryohji F, Ishibashi H:Hepatitis B virus, cigarette smoking and alcohol consumption in the development of hepatocellular carcinoma:a case-control study in Fukuoka,Japan. Int J Cancer 1992,51:509-14.
4 Sherman M:Hepatocellular carcinoma:epidemiology, risk factors, and screening.Semin Liver Dis 2005,25:143-54.
5.Kensler TW, Egner PA, Wang JB, Zhu YR, Zhang BC, Lu PX, Chen JG, Qian GS, Kuang SY, Jackson PE, Gange SJ, Jacobson LP, Muoz A,Groopman JD: Chemoprevention of hepatocellular carcinoma in aflatoxin endemic areas. Gastroenterology 2004,127(5 Suppll):S310-8.
6. Dean M, Fojo T,Bates S. Tumourgkm cells and drug～esistanee[J].Nat REV Cancer,2005,5:275-284.
7 杜珍武 邸军 盛春华 王茜 金正贤 张玉成 张桂珍 甲胎蛋白在肝癌细胞株的癌干细胞中的表达 中国老年学杂志 2008年10月第28卷1896
8 Singh SK, Clarke ID, Terasaki M, al. Identification of a cancer stem cellin human braintumors[J]. CancerRes,2003,63(18):5821-5828.
9.Shmelkov SV, Jun L, St Clair R, el al. Ahemative promoters regulate transcription of the gene that encodes stem cell surface protein AC133 [J]. Blood,2004,103(6): 2055－2061.
10 Peichev M, Naiyer AJ, Pereira D, et al. Expression of VEGFR and AC 133 by circulating human CD34(+)cells identifies population of functional endothelial precursors[J]. Blood,2000 95(3):952-958.
11. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hemutopoielie tell. Nat Med,1997,3(7):730-737. 12. Al-Hajj M,Wicha M S, Benito-Hernandez A,Morrison S J clarke M F. Prospective identification of tumorigenic breast cancer cells [J]. Proc Natl Acad Sci U S A,2003, 100:3983-3988.
13. Singh S K, Hawkins C, Clarke I D, Squire J A, Bayani J, Hide T, etal. Identification of human brain tumour initiating cells [J]. Nature,2004,432: 396-401.
14. Kim C F, Jackson E I, Woolfenden A E, Lawrence S, Babar I, Vogel S, etal. Identification of brOnchiOalveOlar stem cells in normal lung and lung cancer[J]. Cell,2005,121:823-835.
15. Patrawala I, Calhoun T, schneider-BrOussard R, LiH, Bhatia B, Tang S, etal. Highly purified CD44+ prostate cancer cells from xenograft hum an tumors are enriched in turn origenic and metastatic progenitor cells[J]. Oneogene,2006,25: 1696-1708.
16. Suetsugu A, Nagaki M, Aoki H, Motohashi T, Kunisada T,Moriwaki H. Characterization Of CD 133+ hepatOcellular carcinoma cells as cancer stem/ progenitor cells[J]. Biochem Biophys Res Comm un,2006,351:820-824.
17. Chau-TingYeh, Chia-JungKuo, Ming-WeiLai, Tse-ChingChen, Chun-YenLin, Ta-SenYeh and Wei-ChenLee CD133-positive hepatocellular carcinoma in an area endemic for hepatitis B virus infection. BMC Cancer 2009,9:324。
18.张新.CD133在人肝细胞癌中的表达特点及其同患者临床病理因素的关系第二军医大学 博士毕业论文。
1. S Sell. Stem cell origin of cancer and differentiation therapy [JJ.Crit Rev Hematol,2004,51:1-28.
2. Hewitt H B,studies of the dissemination and quantitative transplantation of a lymphocytic leukaemia of CBA mice[J].Br J Cancer,195812:378-401.
3. Park C H, Bergsagel D E, McCulloch E A. Mouse myeloma tumor stem cells: a primary cell culture assay [J]. J Natl Cancer Inst,1971,46:411-422.
4. Hamburger A W, Salmon S E. Primary bioassay of human tumor stem cells[J]. Science,1977,197:461-463.
5. Derkinderen D J, Boxma O J,Koten J W, Den Otter W.Stochastic theory of oncogenesis [J].Anticancer Res,1990,10(2B):497-504.
6. Zajicek G.On the relevant model for human cancer[J].Med Hypotheses,1981,7:1139-1146.
7. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hemutopoielie tell. Nat Med,1997,3(7):730-737.
8. ReyaT, Morrison sJ, Clalke MF, et al. Stem cells, Cancer, and Cancer stem ceils[J]. Nature,2001,414(6859):105-511
9. Pardal R, Clarke MF, Morrison sJ. Applying the principles of stem-cell biology to cancer. Nat Rev Cancer,2003,3(12):895～902.
10. Tsai RY. A molecular view of stem cell and cancer cell selfrenewal. Int J Biochem Cell Biol 2004,36:684-694.
11. Serakinci N, Guldberg P, Burns JS, et al. Adult human mesenchymal sterm cell as a target for neoplastic transformation. Oncogene,2004,23(29):5095.5098.
12. Domen J, GandyKL, Weissman IL. Systemic overexpress of bcl-2 in the hematopoietic system protects transgenic mice from the connsequences of lethal irradiation. Blood,1998。91:2272-2282.
13. Zhou S.Schuetz JD, Bunting KD, et al. The ABC transporter bcrpl/abcg2 is expressed in a wide variety of stem cells and is amolecular determinant of the side-population phenotype. Nature,2001,7:1028.1034.
14. Passegue E, Jamieson C H, Ailles L E, Weissman I L.Normal and leukemic hematopoiesis:are leukemias a stem cell disorder or a reacquisition of stem cell characteristics[J]?Proc Natl Acad Sci USA,2003,100(Suppl 1):11842-11849.
15. Matsui W, Huff CA, Wang Q, et al. Characterization of clonogenic multiple myeloma cells[J]. Blood,2004,103(6):2332-2336.
16. Al-Hajj M,Wicha M S, Benito-Hernandez A,Morrison S J clarke M F. Prospective identification of tumorigenic breast cancer cells [J]. Proc Natl Acad Sci U S A, 2003,100:3983-3988.
17. DontuG, WicAD, Ha MS. Breast cancer, stem/progenitor ceils and the estrogen receptor Trends Endocrinol Metah,2004,15(5):193.197.
18. Singh SK, Clarke ID, Hide T, Dirks P B. Cancer stem cells in nervous system tumors[J]. Oncogene,2004,23:7267-7273.
19. Kim C F, Jackson E L, Woolfenden A E, Lawrence S, Babar I Vogel S, et al. Identification of bronchioalveolar stem cells in normal lung and lung cancer[J]. Cell,2005,121:823-835.
20. Jackson EL, willis N, Mercer K,et al. Analysis of lung tumor initiation and progression using conditional expression of oncogenic K-ras[J]. Genes 2001 15(24):3243-3248.
21. Kim CFB,Jackson,EL,Woolfenden,AE,et al..Identification of bronchioalveolar stem cells in normal lung and lung cancer[J].Cell,2005,121(6):823-835.
22. Lamprecht S A,Lipkin M.Migrating colonic crypt epithelial cells:primary targets for transformation[J].Carcinogenesis,2002,23(11):1777-1780
23. Tatematsu M Tsukamoto T Inada K. Stem cells and gastric cancer:role of gastric and intestinal mixed intestinal metaplasia[J].Cancer Sci,2003,94(2):135-141 o
24. Seigel G M, Campbell I M, Narayan M, Gonzalez-Fernandez F. Cancer stem cell characteristics in retinoblastoma[J]. Mol Vis,2005,11:729-737.
25. Fang D, Nguyen T K, Leishear K, Finko R, Kulp A N, Hotz S, et al. A tumorigenic subpopulation with stem cell properties in melanomas[J]. Cancer Res,2005,65:9328-9337.
26. Li C, Heidt D G, Dalerba P, Burant C F, Zhang I, Adsay V, et al. Identification of pancreatic cancer stem cells[J]. Cancer Res,2007,67:1030-1037.
27. Parkin DM, Bray F, Ferlay J, Pisani P:Estimating the world cancer burden: Globocan 2000. Int J Cancer 2001,94:153-6.
28. Hussain SZ, Sneddon T, Tan X, et al. Wnt impacts growth and differentiation in ex vivo liver development[J]. Exp Cell Res,2004,292(1):157-169.
29. Hixson DC,Faris RA,Thompson NL,et al. An antigenic portrait of the liver during carcinogenesis[J].Pathobiology,1990,58(2):65-67.
30. Lowes KN,Brenan BA,Yeoh GC,et al,Oval cell numbers in human chronic liver diseases are directly related to disease severity[J].Am J Pathol,1999,154(2):537-541.
31. Rountree CB, Senadheera S.Mato JM, et al, Expansion of liver cancer stem cells during aging in methionine adenosyltransferase 1 A_deficient mice. Hepatology, 2008,47:1288-1297.
32. Zhang F, Chen XP, Zhang W, et al. Combined hepatocellular cholangiocarcinom originating from hepatic progenito cells: ImmunohistoehemicaI and doublefluorescence immurtostaining evidence. Histopathology,2008,52:224-232.
33. Yang ZF, Ho DV, Ng MN, et at. Significance of CD90+ Cancer Stem Cells in Human Liver Cancer. Cancer Cell,2008,13:153-166.
34. Yang ZF, Ngai P, Ho DW.et al. Identification of local and circulating cancer stem cells in human liver cancer. Hepatology,2008,47:919-928.
35..杜珍武 邸军 盛春华 王茜 金正贤 张玉成 张桂珍 甲胎蛋白在肝癌细胞株的癌干细胞中的表达 中国老年学杂志 2008年10月第28卷1896
36.Singh SK, Clarke ID, Terasaki M, al. Identification of a cancer stem cellin human braintumors[J]. CancerRes,2003,63(18):5821-5828.
37.Shmelkov SV, Jun L, St Clair R, el al. Ahemative promoters regulate transcription of the gene that encodes stem cell surface protein AC133 [J]. Blood,2004, 103(6):2055-2061.
38 Peichev M, Naiyer AJ, Pereira D, et al. Expression of VEGFR and AC 133 by circulating human CD34(+)cells identifies population of functional endothelial precursors[J]. Blood,2000 95(3):952-958.
39 Mas,Chan KW,Hu L,et al.Identification and characterization of tumorigenic liver cancer stem/progenitor cells[J].Gastroenterology,2007 132(7):2542-2556.
40.Yin S,Li J,Hu C,et al.CD133 positive hepatocellular carcinoma cells possess high capacity for tumorigenicity[J].Int J Cancer,2007,120(7):1444-1450
2.Perz JF, Armstrong GL, Farrington LA, Hutin YJ, Bell BP:The contributions of hepatitis B virus and hepatitis C virus infections to cirrhosis and primary liver cancer worldwide. J Hepatol 2006,45:529-38.
3.Tanaka K, Hirohata T, Takeshita S, Hirohata I, Koga S, Sugimachi K,Kanematsu T, Ohryohji F, Ishibashi H:Hepatitis B virus, cigarette smoking and alcohol consumption in the development of hepatocellular carcinoma:a case-control study in Fukuoka,Japan. Int J Cancer 1992,51:509-14.
4 Sherman M:Hepatocellular carcinoma:epidemiology, risk factors, and screening.Semin Liver Dis 2005,25:143-54.
5.Kensler TW, Egner PA, Wang JB, Zhu YR, Zhang BC, Lu PX, Chen JG, Qian GS, Kuang SY, Jackson PE, Gange SJ, Jacobson LP, Muoz A,Groopman JD: Chemoprevention of hepatocellular carcinoma in aflatoxin endemic areas. Gastroenterology 2004,127(5 Suppll):S310-8.
6. Dean M, Fojo T,Bates S. Tumourgkm cells and drug～esistanee[J].Nat REV Cancer,2005,5:275-284.
7 杜珍武 邸军 盛春华 王茜 金正贤 张玉成 张桂珍 甲胎蛋白在肝癌细胞株的癌干细胞中的表达 中国老年学杂志 2008年10月第28卷1896
8 Singh SK, Clarke ID, Terasaki M, al. Identification of a cancer stem cellin human braintumors[J]. CancerRes,2003,63(18):5821-5828.
9.Shmelkov SV, Jun L, St Clair R, el al. Ahemative promoters regulate transcription of the gene that encodes stem cell surface protein AC133 [J]. Blood,2004,103(6): 2055－2061.
10 Peichev M, Naiyer AJ, Pereira D, et al. Expression of VEGFR and AC 133 by circulating human CD34(+)cells identifies population of functional endothelial precursors[J]. Blood,2000 95(3):952-958.
11. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hemutopoielie tell. Nat Med,1997,3(7):730-737. 12. Al-Hajj M,Wicha M S, Benito-Hernandez A,Morrison S J clarke M F. Prospective identification of tumorigenic breast cancer cells [J]. Proc Natl Acad Sci U S A,2003, 100:3983-3988.
13. Singh S K, Hawkins C, Clarke I D, Squire J A, Bayani J, Hide T, etal. Identification of human brain tumour initiating cells [J]. Nature,2004,432: 396-401.
14. Kim C F, Jackson E I, Woolfenden A E, Lawrence S, Babar I, Vogel S, etal. Identification of brOnchiOalveOlar stem cells in normal lung and lung cancer[J]. Cell,2005,121:823-835.
15. Patrawala I, Calhoun T, schneider-BrOussard R, LiH, Bhatia B, Tang S, etal. Highly purified CD44+ prostate cancer cells from xenograft hum an tumors are enriched in turn origenic and metastatic progenitor cells[J]. Oneogene,2006,25: 1696-1708.
16. Suetsugu A, Nagaki M, Aoki H, Motohashi T, Kunisada T,Moriwaki H. Characterization Of CD 133+ hepatOcellular carcinoma cells as cancer stem/ progenitor cells[J]. Biochem Biophys Res Comm un,2006,351:820-824.
17. Chau-TingYeh, Chia-JungKuo, Ming-WeiLai, Tse-ChingChen, Chun-YenLin, Ta-SenYeh and Wei-ChenLee CD133-positive hepatocellular carcinoma in an area endemic for hepatitis B virus infection. BMC Cancer 2009,9:324。
18.张新.CD133在人肝细胞癌中的表达特点及其同患者临床病理因素的关系第二军医大学 博士毕业论文。
1. S Sell. Stem cell origin of cancer and differentiation therapy [JJ.Crit Rev Hematol,2004,51:1-28.
2. Hewitt H B,studies of the dissemination and quantitative transplantation of a lymphocytic leukaemia of CBA mice[J].Br J Cancer,195812:378-401.
3. Park C H, Bergsagel D E, McCulloch E A. Mouse myeloma tumor stem cells: a primary cell culture assay [J]. J Natl Cancer Inst,1971,46:411-422.
4. Hamburger A W, Salmon S E. Primary bioassay of human tumor stem cells[J]. Science,1977,197:461-463.
5. Derkinderen D J, Boxma O J,Koten J W, Den Otter W.Stochastic theory of oncogenesis [J].Anticancer Res,1990,10(2B):497-504.
6. Zajicek G.On the relevant model for human cancer[J].Med Hypotheses,1981,7:1139-1146.
7. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hemutopoielie tell. Nat Med,1997,3(7):730-737.
8. ReyaT, Morrison sJ, Clalke MF, et al. Stem cells, Cancer, and Cancer stem ceils[J]. Nature,2001,414(6859):105-511
9. Pardal R, Clarke MF, Morrison sJ. Applying the principles of stem-cell biology to cancer. Nat Rev Cancer,2003,3(12):895～902.
10. Tsai RY. A molecular view of stem cell and cancer cell selfrenewal. Int J Biochem Cell Biol 2004,36:684-694.
11. Serakinci N, Guldberg P, Burns JS, et al. Adult human mesenchymal sterm cell as a target for neoplastic transformation. Oncogene,2004,23(29):5095.5098.
12. Domen J, GandyKL, Weissman IL. Systemic overexpress of bcl-2 in the hematopoietic system protects transgenic mice from the connsequences of lethal irradiation. Blood,1998。91:2272-2282.
13. Zhou S.Schuetz JD, Bunting KD, et al. The ABC transporter bcrpl/abcg2 is expressed in a wide variety of stem cells and is amolecular determinant of the side-population phenotype. Nature,2001,7:1028.1034.
14. Passegue E, Jamieson C H, Ailles L E, Weissman I L.Normal and leukemic hematopoiesis:are leukemias a stem cell disorder or a reacquisition of stem cell characteristics[J]?Proc Natl Acad Sci USA,2003,100(Suppl 1):11842-11849.
15. Matsui W, Huff CA, Wang Q, et al. Characterization of clonogenic multiple myeloma cells[J]. Blood,2004,103(6):2332-2336.
16. Al-Hajj M,Wicha M S, Benito-Hernandez A,Morrison S J clarke M F. Prospective identification of tumorigenic breast cancer cells [J]. Proc Natl Acad Sci U S A, 2003,100:3983-3988.
17. DontuG, WicAD, Ha MS. Breast cancer, stem/progenitor ceils and the estrogen receptor Trends Endocrinol Metah,2004,15(5):193.197.
18. Singh SK, Clarke ID, Hide T, Dirks P B. Cancer stem cells in nervous system tumors[J]. Oncogene,2004,23:7267-7273.
19. Kim C F, Jackson E L, Woolfenden A E, Lawrence S, Babar I Vogel S, et al. Identification of bronchioalveolar stem cells in normal lung and lung cancer[J]. Cell,2005,121:823-835.
20. Jackson EL, willis N, Mercer K,et al. Analysis of lung tumor initiation and progression using conditional expression of oncogenic K-ras[J]. Genes 2001 15(24):3243-3248.
21. Kim CFB,Jackson,EL,Woolfenden,AE,et al..Identification of bronchioalveolar stem cells in normal lung and lung cancer[J].Cell,2005,121(6):823-835.
22. Lamprecht S A,Lipkin M.Migrating colonic crypt epithelial cells:primary targets for transformation[J].Carcinogenesis,2002,23(11):1777-1780
23. Tatematsu M Tsukamoto T Inada K. Stem cells and gastric cancer:role of gastric and intestinal mixed intestinal metaplasia[J].Cancer Sci,2003,94(2):135-141 o
24. Seigel G M, Campbell I M, Narayan M, Gonzalez-Fernandez F. Cancer stem cell characteristics in retinoblastoma[J]. Mol Vis,2005,11:729-737.
25. Fang D, Nguyen T K, Leishear K, Finko R, Kulp A N, Hotz S, et al. A tumorigenic subpopulation with stem cell properties in melanomas[J]. Cancer Res,2005,65:9328-9337.
26. Li C, Heidt D G, Dalerba P, Burant C F, Zhang I, Adsay V, et al. Identification of pancreatic cancer stem cells[J]. Cancer Res,2007,67:1030-1037.
27. Parkin DM, Bray F, Ferlay J, Pisani P:Estimating the world cancer burden: Globocan 2000. Int J Cancer 2001,94:153-6.
28. Hussain SZ, Sneddon T, Tan X, et al. Wnt impacts growth and differentiation in ex vivo liver development[J]. Exp Cell Res,2004,292(1):157-169.
29. Hixson DC,Faris RA,Thompson NL,et al. An antigenic portrait of the liver during carcinogenesis[J].Pathobiology,1990,58(2):65-67.
30. Lowes KN,Brenan BA,Yeoh GC,et al,Oval cell numbers in human chronic liver diseases are directly related to disease severity[J].Am J Pathol,1999,154(2):537-541.
31. Rountree CB, Senadheera S.Mato JM, et al, Expansion of liver cancer stem cells during aging in methionine adenosyltransferase 1 A_deficient mice. Hepatology, 2008,47:1288-1297.
32. Zhang F, Chen XP, Zhang W, et al. Combined hepatocellular cholangiocarcinom originating from hepatic progenito cells: ImmunohistoehemicaI and doublefluorescence immurtostaining evidence. Histopathology,2008,52:224-232.
33. Yang ZF, Ho DV, Ng MN, et at. Significance of CD90+ Cancer Stem Cells in Human Liver Cancer. Cancer Cell,2008,13:153-166.
34. Yang ZF, Ngai P, Ho DW.et al. Identification of local and circulating cancer stem cells in human liver cancer. Hepatology,2008,47:919-928.
35..杜珍武 邸军 盛春华 王茜 金正贤 张玉成 张桂珍 甲胎蛋白在肝癌细胞株的癌干细胞中的表达 中国老年学杂志 2008年10月第28卷1896
36.Singh SK, Clarke ID, Terasaki M, al. Identification of a cancer stem cellin human braintumors[J]. CancerRes,2003,63(18):5821-5828.
37.Shmelkov SV, Jun L, St Clair R, el al. Ahemative promoters regulate transcription of the gene that encodes stem cell surface protein AC133 [J]. Blood,2004, 103(6):2055-2061.
38 Peichev M, Naiyer AJ, Pereira D, et al. Expression of VEGFR and AC 133 by circulating human CD34(+)cells identifies population of functional endothelial precursors[J]. Blood,2000 95(3):952-958.
39 Mas,Chan KW,Hu L,et al.Identification and characterization of tumorigenic liver cancer stem/progenitor cells[J].Gastroenterology,2007 132(7):2542-2556.
40.Yin S,Li J,Hu C,et al.CD133 positive hepatocellular carcinoma cells possess high capacity for tumorigenicity[J].Int J Cancer,2007,120(7):1444-1450