同源盒基因A13下调对小细胞肺癌细胞增殖、周期、凋亡和耐药性的影响

The effects of HOXA13 knockdown on cell proliferation,cell cycle,apoptosis,and chemo-resistance of small cell lung cancer

目的探讨小细胞肺癌细胞同源盒基因A13(HOXA13基因)的表达及其下调前后对小细胞肺癌细胞株H146和H446增殖、细胞周期、凋亡以及耐药性的影响。方法利用实时荧光定量PCR(RT-qPCR)法和Western blot法检测小细胞肺癌细胞株H69、H69AR、H1688、H146、H446及人支气管上皮细胞16HBE中HOXA13mRNA和蛋白的表达水平;应用RNAi技术下调HOXA13基因高表达细胞株H146和H446中HOXA13基因表达水平,采用CCK8法和流式细胞术检测HOXA13基因细胞增殖、周期、凋亡及耐药性变化。结果与16HBE细胞相比H69、H1688、H146、H446四种细胞中HOXA13基因高表达;而与H69、H1688细胞相比,H146、H446细胞中HOXA13基因分别相对高表达;下调H146、H446细胞HOXA13基因表达后,细胞增殖受到抑制,细胞周期S期阻滞,细胞凋亡率和药物敏感性增强。结论下调HOXA13基因可抑制小细胞肺癌细胞增殖和促进细胞凋亡,并增强细胞对化疗药物(阿霉素、顺铂和足叶乙甙)的敏感性,提示HOXA13基因可能在小细胞肺癌发生发展及耐药性的产生过程中发挥重要作用。

Objective To investigate the expression of HOXA13 in small cell lung cancer cell lines and the effects of HOXA13 knockdown on cell proliferation,cell cycle,apoptosis and chemo-resistance in H146 and H446 cells. Methods Based on previous microarray results,the expression of HOXA13 at the mRNA and protein level was detected in small cell lung cancer cell lines and the 16 HBE cell line using RT-qPCR and Western blotting.RNAi was used to knockdown HOXA13 in both H146 and H446 cells.The effects of HOXA13 knockdown on cell proliferation,cell cycle,apoptosis,and chemo-resistance were determined.Effects on cell proliferation and chemo-resistance were determined using the Cell Counting Kit-8,and effects on the cell cycle and apoptosis were determined using flow cytometry Results HOXA13 was more highly expressed in H69,H1688,H146,and H446 cells than in 16 HBE cells（P）.HOXA13 was more highly expressed in H446（t=8.939,P=0.0009）and H146 cells（t=9.006,P=0.0008）than in H69 and H1688 cells.HOXA13 knockdown in H146 and H446 cells may lead to the inhibition of cell proliferation,disorder of the cell cycle,and promotion of cell apoptosis and chemo-sensitivity. Conclusion HOXA13 knockdown may inhibit cell proliferation,promote cell apoptosis,and enhance chemo-sensitivity to chemotherapeutic drugs.This suggests that HOXA13 may play a key role in the pathogenesis and chemo-resistance of small cell lung cancer.