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目的:探讨喉癌Hep-2细胞株经流式分选后的非侧群细胞经化疗药物顺铂诱导转变为干细胞样肿瘤细胞的可能机制。方法:喉癌Hep-2细胞株经流式细胞仪荧光活化细胞分选系统分选后,获得的非侧群分别加入头颈部常用化疗药物——顺铂(实验组)和生理盐水(对照组)培养48h,再次上流式细胞仪检测两组侧群细胞比例;并通过RT-PCR及Western blot分别在mRNA和蛋白水平检测两组细胞的干细胞样标记β-catenin、notch-1的基因和蛋白的表达情况。结果:实验组非侧群细胞经顺铂(终浓度1μg/ml)作用48h后,侧群细胞比例为(17.16±0.18)%,对照组则为(10.05±1.20)%,两组差异有统计学意义(t=5.844,P<0.01)。荧光定量RT-PCR示经顺铂处理实验组的β-catenin、notch-1mRNA较对照组显著增多;Western blot示实验组细胞中β-catenin、notch-1蛋白表达量较对照组显著增加(t=5.155,P=0.031;t=5.977,P=0.004)。结论:非侧群细胞经化疗药物顺铂诱导后可分化为干细胞样肿瘤细胞。其可能机制是通过wnt/β-catenin、notch转导通路异常诱导非侧群细胞转化为干细胞样肿瘤细胞。
OBJECTIVE: To investigate the possible mechanism of NSCLC-induced cisplatin-induced transformation of non-side population cells into stem-like tumor cells by flow-sorting of Hep-2 cells. Methods: After laryngeal cancer Hep-2 cells were sorted by flow cytometry fluorescence activated cell sorting system, the non-side groups were added with cisplatin (experimental group) and normal saline (control group) Group) were cultured for 48h. The proportion of side population cells in each group was detected by flow cytometry again. The mRNA and protein levels of β-catenin and notch-1 were detected by RT-PCR and Western blot respectively Protein expression. Results: After treated with cisplatin (1μg / ml) for 48h, the proportion of lateral colony cells was (17.16 ± 0.18)% in control group and (10.05 ± 1.20)% in control group Significance (t = 5.844, P <0.01). The expression ofβ-catenin and notch-1 mRNA in the experimental group was significantly higher than that in the control group by Western blot = 5.155, P = 0.031; t = 5.977, P = 0.004). Conclusion: Non-side population cells can differentiate into stem cell-like tumor cells induced by cisplatin. The possible mechanism is that abnormal transduction pathways of wnt / β-catenin and notch induce non-side population cells to transform into stem cell-like tumor cells.