阿霉素联合紫杉醇调节MAPK/PKC诱导肺癌细胞凋亡的机制研究

Mechanisms of MAPK/PKC Induced Apoptosis by Doxorubicin Combined with Paclitaxel in Human Lung Cancer Cells

目的探讨阿霉素联合紫杉醇调节MAPK/PKC诱导不同种类肺癌细胞凋亡的效果及其机制。方法选取人小细胞肺癌细胞株H446与非小细胞肺癌细胞株A549为实验组,以正常人肺上皮细胞(BEAs-2B)为对照组。CCK-8法检测细胞存活率;检测细胞凋亡蛋白Caspase 3活性;Western blot检测磷酸化MAPK和PKC的蛋白表达水平。同时选取345例肺癌患者,随机接受30天顺铂化疗(158例)和阿霉素联合紫杉醇化疗(187例),检测患者血清MAPK和PKC表达水平,并评价化疗疗效。结果 48 h后,给予阿霉素联合紫杉醇培养的肺癌细胞存活率与对照组相比显著降低,Caspase 3活性显著升高,差异有统计学意义(P<0.05),说明给药后肺癌细胞凋亡明显;与对照组相比,治疗组磷酸化p-p38和p-JNK蛋白表达均明显增加,磷酸化p-PKC蛋白表达明显下降,差异有统计学意义(P<0.05)。阿霉素联合紫杉醇的疗效显著高于单用顺铂,患者血清MAPK水平显著上升,PKC水平显著下降,差异均有统计学意义(P<0.05)。结论阿霉素联合紫杉醇可显著降低肺癌细胞的存活率,促进肺癌细胞凋亡,MAPK/PKC信号通路在此过程中起到重要作用。

Objective： To investigate the mechanism of apoptosis in human lung cancer cells induced by Doxorubicin combined with Paclitaxel. Methods Human small cell lung cancer cell line H446 and non-small cell lung cancer cell line A549 were selected as the experimental group, and the normal human lung epithelial BEAs-2B cell was selected as the control group. CCK-8 assay was used to detect cell viability. Active Caspase 3 was detected by Caspase 3 assay. The expression of phosphorylated MAPK and PKC protein in lung cancer cell lines was detected by Western blot. 345 patients with lung cancer were randomized to receive chemotherapy with cisplatin （158 cases） and chemo- therapy with Doxorubicin and Paclitaxel （187 cases） for 30 days, and the curative effect was evaluated. The expression of MAPK and PKC in serum was detected by ELISA method. Result After treatment of Doxorubicin and Paclitaxel for 48 h, the cell viability of lung cancer ceils was significantly decreased （P〈0.05）, and the activity Of Caspase 3 was significantly increased （P〈0.05）. Western blot showed that the ex- pression of p-p38 and p-JNK were significantly increased （P〈0.05）, and the expression of p-PKC in the lung cancer cells was significantly decreased （P 〈 0.05）. The results showed the effects of chemotherapy with Doxornbicin and Paclitaxel were significantly increased compared with Cisplatin. The MAPK level was significantly increased and PKC level was significantly decreased in serum （P〈0.05）. Conclusion Doxorubicin combined with Paclitaxel could significantly reduce the cell viability of lung cancer cells and induce the apoptosis of lung can- cer cells. MAPK / PKC signaling pathway could play an important role in this process.