基于miR⁃34a/HMGB1轴探讨七氟醚对胃癌细胞恶性生物学活性的机制

Exploration of the mechanism of sevoflurane on malignant biological activity of gastric cancer cells based on miR⁃34a/HMGB1 axis

目的基于miR⁃34a/HMGB1轴探究七氟醚对胃癌细胞恶性生物学活性的影响。方法将胃癌细胞分为NC组,SEV⁃1.7%组、SEV⁃3.4%组、SEV⁃5.1%组。miR⁃34a组、SEV⁃5.1%+anti⁃miR⁃34a组、SEV⁃5.1%+pcDNA⁃HMGB1组和SEV⁃5.1%+pcDNA⁃HMGB1+miR⁃34a组。CCK⁃8、Transwell小室法分别检测细胞增殖和侵袭移能力;qRT⁃PCR检测miR⁃34a表达;蛋白质印迹检测细胞中HMGB1蛋白表达;双荧光素酶报告系统实验检测miR⁃34a和HMGB1的靶向关系。结果和GES⁃1细胞系相比,胃癌SGC⁃7901细胞(0.23±0.03)中miR⁃34a表达降低,HMGB1蛋白(0.85±0.10)表达升高(P<0.05)。和NC组相比,SEV⁃1.7%组、SEV⁃3.4%组、SEV⁃5.1%组细胞中miR⁃34a表达均升高,细胞存活率、细胞侵袭数目和HMGB1蛋白表达均降低(P<0.05)。和SEV⁃5.1%组相比,SEV⁃5.1%+anti⁃miR⁃34a组细胞存活率[24 h、48 h分别为(92.46±8.27)%、(96.71±7.52)%]、细胞侵袭数目(126.42±10.84)和细胞中HMGB1蛋白(0.80±0.09)表达均明显增加(P<0.05)。向细胞中转染HMGB1⁃WT时,miR⁃34a组(0.34±0.05)荧光素酶活性明显低于miR⁃NC组(P<0.05)。和SEV⁃5.1%组相比,SEV⁃5.1%+pcDNA⁃HMGB1组细胞存活率[24 h、48 h分别为(90.18±8.06)%、(82.47±7.21)%]、细胞侵袭数目(122.18±10.62)和细胞中HMGB1蛋白(0.81±0.10)表达均明显升高(P<0.05);和SEV⁃5.1%+pcDNA⁃HMGB1组相比,SEV⁃5.1%+pcDNA⁃HMGB1+miR⁃34a组细胞存活率[24 h、48 h分别为(65.33±6.02)%、(28.41±3.26)%]、细胞侵袭数目(68.33±6.09)和细胞中HMGB1蛋白(0.34±0.04)表达均明显降低(P<0.05)。结论七氟醚可通过调节miR⁃34a/HMGB1轴抑制胃癌细胞增殖和侵袭。

Objective To explore the effect of sevoflurane on the malignant biological activity of gastric can⁃cer cells based on the miR⁃34a/HMGB1 axis.Methods The gastric cancer cells were divided into groups including the NC group,SEV⁃1.7%group,SEV⁃3.4%group,and SEV⁃5.1%group.Additionally,there were groups including the miR⁃34a group,SEV⁃5.1%+anti⁃miR⁃34a group,SEV⁃5.1%+pcDNA⁃HMGB1 group,and SEV⁃5.1%+pcDNA⁃HMGB1+miR⁃34a group.Cell proliferation was assessed using the CCK⁃8 assay,while cell invasion and migration capabilities were assessed using the Transwell chamber assay.The expression of miR⁃34a was measured using qRT⁃PCR,and the expression of HMGB1 protein in cells was detected using Western blot analysis.The targeting relation⁃ship between miR⁃34a and HMGB1 was investigated using a dual luciferase reporter system experiment.Results Compared to the GES⁃1 cell line,miR⁃34a expression was downregulated(0.23±0.03)and HMGB1 protein expres⁃sion was upregulated(0.85±0.10)in gastric cancer SGC⁃7901 cells(P<0.05).Compared to the NC group,miR⁃34a expression increased and cell viability,cell invasion,and HMGB1 protein expression decreased in the SEV⁃1.7%group,SEV⁃3.4%group,and SEV⁃5.1%group(P<0.05).Compared to the SEV⁃5.1%group,cell viability(92.46%±8.27%,96.71%±7.52%in 24 h and 48 h,respectively),cell invasion(126.42±10.84),and HMGB1 protein expression(0.80±0.09)significantly increased in the SEV⁃5.1%+anti⁃miR⁃34a group(P<0.05).When trans⁃fecting HMGB1⁃WT into cells,the luciferase activity of the miR⁃34a group(0.34±0.05)was significantly lower than that of the miR⁃NC group(P<0.05).Compared to the SEV⁃5.1%group,cell viability(90.18%±8.06%,82.47%±7.21%in 24 h and 48 h,respectively),cell invasion(122.18±10.62),and HMGB1 protein expression(0.81±0.10)significantly increased in the SEV⁃5.1%+pcDNA⁃HMGB1 group(P<0.05).Compared to the SEV⁃5.1%+pcD⁃NA⁃HMGB1 group,cell viability(65.33%±6.02%,28.41%±3.26%in 24 h and 48 h,respectively),cell invasion(68.33±6.09),and HMGB1 protein expression(0.34±0.04)significantly decreased in the SEV⁃5.1%+pcDNA⁃HMGB1+miR⁃34a group(P<0.05).Conclusion Sevoflurane can inhibit the proliferation and invasion of gastric cancer cells by regulating the miR⁃34a/HMGB1 axis.