转染pro-GRP表达载体通过JAK1/STAT3信号通路诱导小细胞肺癌细胞H446凋亡的实验研究-东南大学学报医学版

转染pro-GRP表达载体通过JAK1/STAT3信号通路诱导小细胞肺癌细胞H446凋亡的实验研究

单位：1. 成都大学医学院, 四川成都 610106;
2. 成都市第五人民医院肿瘤科, 四川成都 611130

关键词：小细胞肺癌胃泌素释放肽前体 Janus激酶1/信号转导子和转录激活子3 凋亡增殖

分类号：R734.2

出版年·卷·期（页码）：2019·38·第四期（661-666）

摘要：

目的：探讨胃泌素释放肽前体（pro-GRP）对小细胞肺癌细胞H446增殖的影响及其分子机制。方法：培养小细胞肺癌细胞H446，将其分为阴性对照组（转染空白质粒）、pro-GRP过表达组（转染pro-GRP表达质粒）、pro-GRP过表达+AG490组（转染pro-GRP表达质粒并用STAT3抑制剂AG490处理）。处理24 h后测定细胞的增殖活力以及线粒体凋亡基因Bcl-2、Bax、Caspase-3和JAK1/STAT3信号通路分子的表达量。结果：pro-GRP、p-JAK1、p-STAT3的蛋白表达量pro-GRP过表达组高于阴性对照组，差异均有统计学意义（P<0.05）。细胞活力检测450 nm处的吸光度、Bcl-2的蛋白表达量pro-GRP过表达组高于阴性对照组，差异均有统计学意义（P<0.05）；Bax、Caspase-3的蛋白表达量pro-GRP过表达组低于阴性对照组，差异均有统计学意义（P<0.05）。细胞活力检测450 nm处的吸光度、Bcl-2的蛋白表达量pro-GRP过表达+AG490组低于pro-GRP过表达组，差异均有统计学意义（P<0.05）；Bax、Caspase-3的蛋白表达量pro-GRP过表达+AG490组高于pro-GRP过表达组，差异均有统计学意义（P<0.05）。结论：转染pro-GRP表达载体能够通过JAK1/STAT3信号通路抑制线粒体途径细胞凋亡并促进小细胞肺癌细胞H446的增殖。

Objective: To investigate the effect of pro-GRP on the proliferation of small cell lung cancer cell H446 and its molecular mechanism. Methods: Small cell lung cancer cells H446 were cultured and divided into negative control group (transfected with blank plasmid), pro-GRP overexpression group (transfected with pro-GRP expression plasmid), pro-GRP overexpression+AG490 group (transfected with pro-GRP expression plasmid and STAT3 inhibitor AG490). After 24 hours of treatment, the cell proliferation activity and the expression of mitochondrial apoptotic genes Bcl-2, Bax, Caspase-3 and JAK1/STAT3 signaling pathway were determined. Results: The protein expression levels of pro-GRP, p-JAK1 and p-STAT3 in the pro-GRP overexpression group were significantly higher than those in the negative control group (all P<0.05). The light absorption value and the protein expression level of Bcl-2 in the pro-GRP overexpression group at 450 nm were higher than those in the negative control group, and the protein expression levels of Bax and Caspase-3 were significantly lower than those in the negative control group (all P<0.05). The light absorption value and the protein expression level of Bcl-2 in the pro-GRP overexpression+AG490 group at 450 nm were lower than those in the pro-GRP overexpression group (all P<0.05). The protein expression levels of Bax and Caspase-3 in the pro-GRP overexpression +AG490 group were significantly higher than those in the pro-GRP overexpression group (P<0.05).Conclusion: Transfection of pro-GRP expression vector can inhibit mitochondrial pathway apoptosis and promote the proliferation of small cell lung cancer cell line H446 through JAK1/STAT3 signaling pathway.

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