Initial Study Of Cd133 As One Of Surface Markers In Hepatocellular Carcinoma Stem Cell

Part 1:The Property of CD133+ Cell in Hepatic Cell Lines in VitroObjective:To isolate CD133+ cell in hepatic cell lines SMMC7721and Bel-7402 by magnetic activated cell sorting (MACS) system, and to observe its biological characteristics in vitro.Methods : The expression of stem cell marker CD133 in hepatic cell lines SMMC7721 and Bel-7402,and the cell cycle was detected by flow cytometry. Magnetic cell sorting was used to isolate CD133+and CD133-cells.The difference in morphology, differentiation and proliferation between CD133+ and CD133- in vitro were observed and analyzed by one-way ANOVA and u test.Results :The percentages of CD133+ cell in SMMC7721 and Bel-7402 cell lines were 0.7%-1.0% and 1.7%-8.9%, respectively. The percentages of CD133+ cell in G0/G1 phase in the two cell lines were 85.3% and 89.4%, which were significantly higher than CD133-cells and unsorted cells(F=14.49 , P<0.05). The in vitro proliferation capability of CD133+ cells was greater than that of CD133-and unsorted cells,especially during day 1-3and day 5-7(F=49.32,784.04,89.91,152.83,P<0.05). During the cultivation,the proportion of the CD133+ cells decreased as time passed by,and the proportion of CD133-cells was nearly the same as unsorted cells on day 15(U=0.271,P<0.05). Conclusions:CD133+ cells have strong capability of differentiation and proliferation in SMMC7721 and Bel-7402 cell lines in vitro, which have biological characteristic of cancer stem cell.Part 2: Expression and clinical significance of CD133 in hepatocellular carcinoma tissuesObjective:To study the expression and clinical significance of cancer stem cell marker CD133 in hepatocellular carcinoma tissues.Methods:The expression of CD133 mRNAs which was extracted from human hepatocellular carcinoma tissues(HCT) and paracarcinoma tissues(PT) of 25 patients were detected by RT-PCR. The relationships between CD133 and clinicopathological index were analysed.Results:The positive rate of the CD133 mRNA expression in HCT was 72%, while it was 4% in PT(P<0.01).The expression of CD133 mRNA was correlated with tumor size and the expression of AFP(P<0.05).There was no evidence indicating that was relative with sex, age and tumor differentiation (P>0.05).Conclusion:The expression of cancer stem cell marker CD133 was related to tumorigenesis of hepatocellular cancer and the expression of AFP. CD133 may be used in diagnosing and judging the prognosis of hepatocellular cancer.