瑞芬太尼通过miR-519d-3p/STAT3对胃癌细胞增殖、凋亡的影响

Remifentanil inhibits proliferation and promotes apoptosis of gastric cancer cells by regulating miR-519d-3p/STAT3 expression

背景研究表明,阿片类药物不仅用于肿瘤手术麻醉及术后镇痛,而且对肿瘤细胞恶性生物学行为具有一定抑制作用.瑞芬太尼作为一种高效阿片受体激动剂,已有报道瑞芬太尼可抑制结肠癌、肝癌、肺腺癌等多种肿瘤细胞增殖、迁移并诱导肿瘤细胞凋亡.但目前瑞芬太尼对胃癌(gastric cancer, GC)细胞增殖和凋亡的影响及其机制尚不清楚.目的研究瑞芬太尼对GC细胞增殖、凋亡的影响及其潜在作用机制.方法培养人GC SGC7901细胞, MTT实验检测不同浓度瑞芬太尼对细胞增殖的影响.构建过表达miR-519d-3p的GC细胞,另构建抑制表达miR-519d-3p和过表达STAT3的细胞并给于瑞芬太尼处理,分别于培养24h、48h和72h时采用MTT法测定细胞增殖活性,培养48 h时采用流式细胞仪测定细胞凋亡情况, qRTPCR和Westernblot实验分别用于检测细胞中miR-519d-3p和STAT3蛋白表达水平,双荧光素酶报告基因实验检验miR-519d-3p是否靶向STAT3.结果瑞芬太尼能够有效抑制GC细胞增殖,诱导细胞凋亡,并促进细胞中miR-519d-3p表达;过表达miR-519d-3p抑制GC细胞增殖并诱导细胞凋亡,而抑制miR-519d-3p表达可逆转瑞芬太尼对GC细胞增殖和凋亡的作用;双荧光素酶报告基因实验证实GC细胞中miR-519d-3p靶向负调控STAT3的活性;过表达STAT3逆转了瑞芬太尼对GC细胞增殖和凋亡的作用.结论瑞芬太尼具有抑制GC细胞增殖并诱导细胞凋亡的作用,其作用机制可能与调控miR-519d-3p/STAT3表达有关.

BACKGROUND Studies have shown that opioids can not only be used for tumor anesthesia and postoperative analgesia, but also have an inhibitory effect on the malignant biological behavior of tumor cells. As a highly efficient opioid receptor agonist, remifentanil has been reported to inhibit the proliferation and migration and induce apoptosis of tumor cells such as colon cancer, liver cancer, and lung adenocarcinoma cells. However, the effect of remifentanil on the proliferation and apoptosis of gastric cancer(GC) cells and the underlying mechanism are still unclear.AIM To study the effect of remifentanil on proliferation and apoptosis of GC cells and the potential mechanism involved. METHODS Human GC SGC7901 and BGC823 cells were cultured, and the effects of different concentrations of remifentanil on cell proliferation were detected by MTT assay. GC cells overexpressing miR-519d-3p, those with low expression of miR-519d-3p, and those overexpressing STAT3 were constructed and treated with remifentanil. Cell proliferation activity was measured by MTT assay at 24 h, 48 h, and 72 h after remifentanil treatment. Apoptosis was detected by flow cytometry at 48 h. qRTPCR and Western blot were used to detect the expression levels of miR-519d-3p and STAT3 in cells, respectively. The dual luciferase reporter assay was used to test whether miR-519d-3p targets STAT3. RESULTS Remifentanil could effectively inhibit the proliferation of GC cells, induce their apoptosis, and promote the expression of miR-519d-3p. Overexpression of miR-519d-3p inhibited the proliferation of GC cells and induced their apoptosis, while inhibition of miR-519d-3p expression reversed the effect of remifentanil on the proliferation and apoptosis of GC cells. Dual luciferase reporter gene assay confirmed that miR-519d-3p negatively regulated STAT3 activity in GC cells. Overexpression of STAT3 reversed the effect of remifentanil on the proliferation and apoptosis of GC cells. CONCLUSION Remifentanil can inhibit the proliferation of GC cells and induce their apoptosis, which may be related to the regulation of mir-519d-3p/STAT3 expression.