肺癌细胞株中EPS8表达下降对顺铂化疗敏感性的影响

发布时间：2019-05-09 05:26

【摘要】：目的探究肺癌细胞株中表皮生长因子受体通路底物8(EPS8)表达下降对顺铂化疗敏感性的影响。方法脂质体转染法将EPS8沉默后并通过实时荧光定量聚合酶链反应(qRT-PCR)检测EPS8在各细胞系的沉默效率。采用Caspase-3/7细胞凋亡实验检测,5μmol顺铂对淋巴母细胞样细胞系(LCLs)和A549肺癌细胞凋亡的影响。Alamar Blue细胞生长抑制实验中采用0.01μmol/L光神霉素A作为EPS8抑制剂单用及联用不同浓度顺铂,Alamar Blue细胞生长抑制实验检测其对癌和非癌细胞系生长抑制的影响。结果与对照组比较,LCLs细胞系中EPS8的沉默导致顺铂干预时细胞存活率升高7.9%,凋亡率下降8.7%。与此相反,顺铂干预导致肺癌细胞存活率下降20.6%。光神霉素A在LCLs细胞系中能够降低EPS8的表达,并与对照组比较,非癌细胞系LCLs在顺铂干预时具有更低Caspase-3/7活性,凋亡率降低。在5种非小细胞肺癌(NSCLC)细胞系中,光神霉素A同样能够导致EPS8表达下降。与LCLs细胞系比较,光神霉素A的干预能够使4种NSCLC细胞和膀胱癌细胞HTB9对顺铂的敏感性增加(P0.05),但NSCLC中存在EGFR突变的H1975细胞对顺铂的敏感性并未出现统计学改变(P0.05)。结论通过核糖核酸(RNA)干扰或者光神霉素A将EPS8表达抑制后,肿瘤细胞对顺铂的敏感性升高,而正常细胞LCLs对其敏感性却出现下降。对EPS8分子机制的进一步研究有望其成为抗肿瘤治疗的新靶点。
[Abstract]:Objective to investigate the effect of decreased expression of epidermal growth factor receptor pathway substrate 8 (EPS8) on chemotherapy sensitivity of cisplatin in lung cancer cell lines. Methods the silencing efficiency of EPS8 in various cell lines was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) after silencing with lipofectamine. Caspase-3/7 cell apoptosis assay was used. The effect of 5 渭 mol DDP on apoptosis of lymphoid cell line (LCLs) and A549 lung cancer cells. Alamar Blue cells were treated with 0.01 渭 mol / L photosensin A as an inhibitor of EPS8 alone and in combination with different concentrations of DDP. The effect of Alamar Blue cell growth inhibition test on the growth inhibition of cancer and non-cancer cell lines was detected. Results compared with the control group, the silencing of EPS8 in LCLs cell line increased the cell survival rate by 7.9% and decreased the apoptosis rate by 8.7%. In contrast, the survival rate of lung cancer cells decreased by 20.6%. Photomycin A could decrease the expression of EPS8 in LCLs cell line. Compared with the control group, the non-cancer cell line LCLs had lower Caspase-3/7 activity and lower apoptosis rate when treated with cisplatin. In five non-small cell lung cancer (NSCLC) cell lines, photoShenmycin A can also lead to the decrease of EPS8 expression. Compared with LCLs cell line, the intervention of photomycin A increased the sensitivity of four NSCLC cells and bladder cancer cell line HTB9 to cisplatin (P0.05). However, there was no significant change in the sensitivity of H 1975 cells with EGFR mutation to DDP in NSCLC (P 0.05). Conclusion after EPS8 expression was inhibited by ribonucleic acid (RNA) interference or photomycin A, the sensitivity of tumor cells to cisplatin increased, while the sensitivity of normal LCLs cells to cisplatin decreased. Further study on the molecular mechanism of EPS8 is expected to be a new target for antitumor therapy.
【作者单位】：广州军区广州总医院呼吸内科;广东药科大学;中国人民解放军第四军医大学;
【分类号】：R734.2

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