乳酸脱氢酶A基因敲除联合丝裂霉素C对前列腺癌细胞凋亡的影响及其机制
Combined effects and mechanism of lactate dehydrogenase A gene knockout and mitomycin C on prostate cancer cell apoptosis
目的:探讨乳酸脱氢酶A(LDHA)基因敲除联合丝裂霉素C(MMC)对前列腺癌细胞株PC3细胞凋亡的影响及其机制。方法免疫印迹法检测LDHA基因敲除PC3细胞(LDHA PC3)的基因敲除效果,细胞侵袭实验检测LDHA PC3细胞与PC3细胞的侵袭力。流式细胞术检测MMC处理和LDHA基因敲除对PC3细胞凋亡的影响,免疫印迹法检测凋亡通路相关蛋白表达变化。结果免疫印迹显示,KOLDHA PC3细胞株3个克隆均确定完整敲除LDHA基因。敲除LDHA基因能抑制PC3细胞的侵袭力,并通过caspase?3/二磷酸腺苷核糖聚合酶(PARP)凋亡通路促进MMC诱导的PC3细胞凋亡。结论 LDHA基因敲除联合MMC可协同作用诱导PC3细胞凋亡。
更多Objective To investigate the effects and mechanism of lactate dehydrogenase A (LDHA)gene knockout combined with mitomycin C(MMC)on apoptosis of prostate cancer cell line PC3. Methods Western blotting was used to validate the LDHA gene knockout PC3 cells(LDHA PC3). Cell invasion assay was used to examine the invasiveness of LDHA PC3 cells and PC3 cells. Flow cytometry was used to investigate the effects of MMC treatment and LDHA gene knockout on PC3 apoptosis. Western blotting was employed to determine the altered expression of proteins related to apoptosis pathway. Results Western blotting validated complete LDHA gene knockout in 3 clones of KOLDHA PC3 cell line. LDHA gene knockout was shown to inhibit the invasiveness of PC3 cells,and to promote MMC?induced PC3 cell apoptosis via the caspase?3/poly ADP?ribose polymerase (PARP) pathway. Conclusion LDHA gene knockout and MMC may work synergistically to induce apoptosis of PC3 cells.
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