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目的：〖HT5"SS〗建立稳定过表达微粒体谷胱甘肽S转移酶1（microsomal glutathione Stransferase 1, 〖STBX〗MGST1〖STBZ〗）基因的肺腺癌SPCA1细胞系，探讨〖STBX〗MGST1〖STBZ〗在肺腺癌中的作用及其机制。〖HT5W〗方法：〖HT5"SS〗重组质粒pcDNA3MGST1和空载体pcDNA3以脂质体介导的方法转染至SPCA1细胞中，经过G418筛选稳转细胞系，标记为pcDNA3MGST1细胞和空载体pcDNA3细胞。实时荧光定量PCR及Western blotting鉴定稳转细胞中〖STBX〗MGST1〖STBZ〗 mRNA和蛋白的表达情况。MTS法检测稳转细胞的活力;流式细胞仪和Western blotting检测H2O2诱导下稳转细胞的凋亡率和下游凋亡相关蛋白水平变化。〖HT5W〗结果：〖HT5"SS〗酶切鉴定和测序结果显示pcDNA3MGST1重组质粒构建成功，并获得具有G418抗性的稳转细胞株。pcDNA3MGST1细胞中〖STBX〗MGST1〖STBZ〗在mRNA和蛋白水平表达均显著升高（P<0.01），且细胞活力明显增加（P<0.05）。H2O2诱导下，pcDNA3MGST1细胞的早期凋亡率明显低于pcDNA3组[（330±0.40）% vs （6.50±0.95）%, P<0.05];pcDNA3MGST1细胞凋亡相关蛋白caspase 9、caspase 3、 PARP表达增多，cleavedcaspase 9、cleavedcaspase 3、cleavedPARP表达明显减少。〖HT5W〗结论：〖HT5"SS〗本研究成功构建了稳定过表达〖STBX〗MGST1〖STBZ〗的SPCA1肺腺癌细胞系，〖STBX〗MGST1〖STBZ〗可能通过调节caspase凋亡通路抑制肺腺癌细胞的凋亡。
Objective: 〖WTBZ〗 To establish a lung adenocarcinoma SPCA1 cell stably expressing microsomal glutathione Stransferase 1 ( 〖STBX〗 MGST1〗 〖STBZ〗) To investigate the role and mechanism of STBX〗 MGST1 STBZ in lung adenocarcinoma. 〖HT5W〗 Methods: 〖WTBZ〗 The recombinant plasmid pcDNA3-MGST1 and empty vector pcDNA3 were transfected into SPCA1 cells by liposomemediated method. After G418 selection, the cells were stably transfected and labeled with pcDNA3 MGST1 cells and empty vector pcDNA3 cells. Real-time quantitative PCR and Western blotting were used to identify the mRNA and protein expression of 〖STBX〗 MGST1 〖STBZ〗 in metastatic cells. MTS method was used to detect the viability of metastatic cells. Flow cytometry and Western blotting were used to detect the apoptotic rate and the downstream apoptosis-related proteins in the stable transfected cells induced by H2O2. 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖WTBZ〗 〖HTZ〗 The mRNA and protein levels of MGST1 and STBZ in pcDNA3MGST1 cells were significantly increased (P <0.01), and the cell viability was significantly increased (P <0.05). The early apoptotic rate of pcDNA3MST1 cells was significantly lower than that of pcDNA3 group (330 ± 0.40)% vs (6.50 ± 0.95)% under the induction of H2O2, The expression of caspase 9, caspase 3 and PARP in pcDNA3-MGST1 cells increased and the expression of cleaved-caspase 9, cleaved-caspase 3 and cleaved-PARP decreased significantly. 〖WTBZ〗 〖HTS〗 〖HTS〗 〖HTS〗 In this study, SPCA1 lung adenocarcinoma cell line stably overexpressed 〖STBX〗 MGST1 〖STBZ〗 was constructed, and 〖STBX〗 MGST1〗 〖STBZ might pass Regulation of caspase apoptosis pathway inhibits apoptosis of lung adenocarcinoma cells.