七氟醚通过lncRNA MEG3调控miR-23a-3p对肺癌细胞增殖和凋亡的影响

Effects of sevoflurane on the regulation of miR-23a-3p on lung cancer cell proliferation and apoptosis through lncRNA MEG3

目的研究七氟醚对肺癌细胞增殖和凋亡的影响及其分子机制。方法采用七氟醚处理肺癌A549细胞,CCK-8检测细胞增殖,流式细胞术评估细胞凋亡,Western blot检测B细胞淋巴瘤2(Bcl-2)蛋白和Bcl-2相关X蛋白(Bax)表达。实时定量PCR(qRT-PCR)检测长链非编码RNA(lncRNA)MEG3和微小RNA(miR)-23a-3p的表达。qRT-PCR分析人正常肺粘膜GES-1细胞和肺癌A549细胞中MEG3和miR-23a-3p表达。生物信息学预测与双荧光素酶活性实验检测肺癌细胞A549中MEG3与miR-23a-3p的靶向关系。在A549细胞中转染si-MEG3或miR-23a-3p,并分别给予七氟醚处理,采用上述方法进行细胞增殖与凋亡检测。结果七氟醚明显降低细胞A549的活力、Bcl-2蛋白水平和miR-23a-3p表达量,显著提高细胞的凋亡率、Bax蛋白表达量和MEG3表达量(P<0.05)。与GES-1细胞比较,A549细胞MEG3表达量明显减少,miR-23a-3p表达量显著增加(P<0.05)。MEG3靶向调控miR-23a-3p的表达。七氟醚通过MEG3抑制miR-23a-3p的表达。敲减MEG3或过表达miR-23a-3p后,七氟醚对A549细胞增殖和Bcl-2蛋白表达的抑制作用,以及对细胞凋亡和Bax蛋白表达的促进作用被逆转。并且,七氟醚对A549细胞miR-23a-3p表达的抑制作用被敲减MEG3所逆转。结论七氟醚通过lncRNA MEG3调控miR-23a-3p的表达,抑制肺癌细胞增殖和诱导细胞凋亡。

Objective To study the effect of sevoflurane on the proliferation and apoptosis of lung cancer cells and the molecular mechanism.Methods Lung cancer A549 cells were treated with sevoflurane.Cell proliferation was measured by cell counting kit 8(CCK-8).Flow cytometry assessed cell apoptosis.Bcl-2 related X protein(Bax)and B-cell lymphoma/leukemia 2(Bcl-2)protein expression were determined by Western blot,real-time quantitative PCR(qRT-PCR)was used to detect long-chain non-coding RNA(lncRNA)MEG3 and microRNA(miR)-23 a-3 p expression.qRT-PCR analysis of MEG3 and miR-23 a-3 p expression in human normal lung mucosa cells GES-1 and lung cancer cells A549.Bioinformatics prediction and dual luciferase activity assay were applied to detect the targeting relationship between MEG3 and miR-23 a-3 p in lung cancer A549 cells.A549 Cells were transfected with si-MEG3 or miR-23 a-3 p and treated with sevoflurane,respectively.Cell proliferation and apoptosis were detected using the methods described above.Results Sevoflurane significantly reduced A549 cell viability,Bcl-2 protein level and miR-23 a-3 p expression,and obviously improved cell apoptosis rate,increased Bax protein expression,and MEG3 expression(P<0.05).Compared with cell GES-1,MEG3 expression in lung cancer A549 cells was dramatically decreased,and miR-23 a-3 p expression was remarkably increased(P<0.05).MEG3 targets the regulation of miR-23 a-3 p expression.Sevoflurane inhibits the expression of miR-23 a-3 p through MEG3.After MEG3 was knocked out or miR-23 a-3 p was overexpressed,the inhibitory effects of sevoflurane on A549 cell proliferation and Bcl-2 protein expression,and the promotion of apoptosis and Bax protein expression were reversed.Moreover,the inhibitory effect of sevoflurane on miR-23 a-3 p expression in A549 cells was reversed by knockdown of MEG3.Conclusion Sevoflurane regulates the expression of miR-23 a-3 p through lncRNA MEG3,inhibits lung cancer cell proliferation and induces apoptosis.