【摘要】 目的探讨塞来昔布对幽门螺杆菌(Helicobacter pylori,H.pylori)感染的胃癌细胞凋亡及炎症反应的影响及机制。方法以终浓度为50、75、100μmol/L的塞来昔布作用于H.pylori感染的人胃癌SGC-7901细胞,MTT法检测作用24、48和72 h的细胞增殖;作用48 h,流式细胞术检测细胞凋亡率,Western blotting检测PCNA、Bcl-2、Bax和p-AKT3蛋白表达,qRT-PCR检测IL-6、IL-8和miR-145表达。双荧光素酶报告基因实验检测miR-145和AKT3的靶向关系。结果不同浓度塞来昔布均可明显抑制SGC-7901细胞增殖,且呈剂量、时间依赖性(P<0.05)。不同浓度塞来昔布均可明显促进细胞凋亡,下调PCNA、Bcl-2、p-AKT3、IL-6和IL-8表达,上调Bax和miR-145表达,呈剂量依赖性(P<0.05)。双荧光素酶报告基因实验结果显示,miR-145和AKT3存在靶向关系。结论塞来昔布可抑制H.pylori感染的胃癌细胞增殖,促进细胞凋亡,降低炎症因子IL-6和IL-8表达,机制可能与塞来昔布引起miR-145表达改变进而调控其靶基因AKT3表达有关。更多还原

【Abstract】 Objective To investigate the effect and mechanism of Celecoxib on apoptosis and inflammatory response of gastric cancer cells infected with Helicobacter pylori(H.pylori). Methods Human gastric cancer SGC-7901 cells were treated with Celecoxib(50, 75, 100 μmol/L). The proliferation of SGC-7901 cells was detected by MTT assay at 24 hours, 48 hours and 72 hours. Cells were treated for 48 hours, the apoptosis rate was detected by flow cytometry, the protein expression of PCNA, Bcl-2, Bax and p-AKT3 was detected by Western blotting, and the expression of IL-6, IL-8 and miR-145 was detected by qRT-PCR. Double luciferase reporter gene assay was used to detect the targeting relationship between miR-145 and AKT3. Results Different concentrations of Celecoxib could significantly inhibit the proliferation of SGC-7901 cells in a dose-and time-dependent manner(P<0.05). Different concentrations of Celecoxib could significantly promote apoptosis, down regulate the expression of PCNA, Bcl-2, p-AKT3, IL-6 and IL-8, and up regulate the expression of Bax and miR-145 in a dose-dependent manner(P<0.05). Double luciferase reporter gene assay showed that there was targeting relationship between miR-145 and AKT3. Conclusion Celecoxib can inhibit the proliferation of H.pylori-infected gastric cancer cells, promote cell apoptosis, and reduce the expression of inflammatory factors IL-6 and IL-8. The mechanism may be related to the change of miR-145 expression caused by Celecoxib to regulate its target genes AKT3 expression.更多还原