顺铂对HepG2细胞增殖、细胞周期及肝癌干细胞标志物的影响

Effects of Cisplatin on HepG2 Cell Proliferation, Cell Cycle and Markers of Liver Cancer Stem Cells

目的:研究顺铂对HepG2细胞增、细胞周期及肝癌干细胞标志物(CD133、ICAM-1和ABCG2)的影响。方法:选取HepG2作为研究对象,分别采用MMT比色法、PI染色法及免疫荧光法检测不同浓度顺铂对其增殖、细胞周期、CD133、ICAM-1和ABCG2表达的影响。结果:每个浓度顺铂作用后均可以显著抑制HepG2细胞增殖力(F=12.23,P=0.004);顺铂对HepG2细胞增殖力的抑制作用和浓度可能与时间成正比。0 mg/L组静息期(G0/G1期)细胞比例为(50.25±0.79)%、2 mg/L组G0/G1期细胞比例为(89.24±0.41)%、4 mg/L组G0/G1期细胞比例为(29.54±3.02)%,2 mg/L组和4 mg/L组分别比0 mg/L组显著上升和下降,差异明显有统计学意义(t=6.53、-3.65,均P<0.05)。0 mg/L组DNA合成期(S期)细胞比例为(47.13±0.74)%、2 mg/L组S期细胞比例为(5.65±0.42)%、4 mg/L组S期细胞比例为(67.46±3.24)%,2 mg/L组和4 mg/L组分别比0 mg/L组显著下降和上升,差异明显有统计学意义(t=-7.35、3.79,均P<0.05)。结果提示2 mg/L组和4 mg/L组顺铂可让HepG2在G0/G1期与S期显著阻滞,差异有统计学意义(P<0.01);顺铂处理后,剩余的HepG2细胞的CD133、ICAM-1和ABCG2呈现高表达水平。结论:HepG2细胞系中会有很少部分肝癌干细胞避开了顺铂的杀灭作用,是造成临床上肝癌反复发作的重要因素之一,临床上应予以重视。

Objective： To study the effect of cisplatin on HepG2 cell proliferation, cell cycle and liver cancer stem cell markers（CD133, ICAM-1 and ABCG2）. Methods： Selecting HepG2 as the research object, we used MMT colorimetric assay to measure the proliferation of cells with different concentrations of cisplatin; we used PI staining to measure the cell cycle after cisplatin treatment; the expression of CD133, ICAM-1 and ABCG2 were measured by immunofluorescence assay. Results： Each concentration of cisplatin could significantly inhibit the proliferation of HepG2 cells（F=12.23, P=0.004）; The inhibitory effect and concentration of cisplatin on the proliferation of HepG2 cells may be proportional to the time. In 0 mg/L group, the proportion of quiescent phase（G0/G1） cells was（50.25±0.79） %, the ratio of G0/G1 phase to G1 phase was（89.24 ±0.41）%, the ratio of 4 mg/L group G0/G1 cells were（29.54 ±3.02）%, 2 mg/L group and 4 mg/L group were significantly higher than 0 mg/L group（t=6.53,-3.65, P〈0.05）. The proportion of DNA in0 mg/L group was（47.13±0.74） %, the proportion of s phase in 2 mg/L group was（5.65±0.42）%, the proportion of s phase in 4 mg/L group was（67.46±3.24）%, the ratio of 2 mg/L group and 4 mg/L group decreased significantly and increased significantly（t=-7.35, 3.79,P〈0.05）. The results showed that cisplatin in 2 mg/L group and 4 mg/L group could significantly block HepG2 in G0/G1 phase and s phase, with significant difference（P〈0.01）. After cisplatin treatment, CD133, ICAM-1 and ABCG2 showed high expression levels in the remaining HepG2 cells. Conclusion： In the HepG2 cell line, there were a small number of liver cancer stem cells to avoid the killing effect of cisplatin, which was one of the important clinical factors of recurrent liver cancer, so we should be paid attention to in clinical.