目的 研究吉马酮通过HBXIP调控PRAS40/p-PRAS4影响胃癌细胞增殖的机制。方法 收集20例行胃癌切除术患者的胃癌组织及配对的癌旁组织，通过免疫组化实验及Western blotting实验检测乙型肝炎X相互作用蛋白（HBXIP）、相对分子质量为4×104的富含脯氨酸蛋白激酶B底物蛋白（PRAS40）、磷酸化PRAS40（p-PRAS40）的表达量；以20例癌组织中HBXIP、PRAS40、p-PRAS40蛋白表达的免疫组织化学染色评分为数据，通过Correlation matrix分析PRAS40、p-PRAS40的表达与HBXIP表达的相关性；在胃癌MGC803、SGC7901细胞中通过转染质粒过表达或沉默HBXIP后（NC组转染空白质粒），MTT实验检测细胞增殖能力的变化，通过流式细胞术及Hoechst 33285染色检测细胞凋亡情况，并通过Westernblotting实验检测HBXIP、PRAS40、p-PRAS40的蛋白表达量；以0（溶剂对照组）、50、100、150、200、250 μmol·L^-1吉马酮处理胃癌MGC803、SGC7901细胞48 h，MTT实验检测细胞增殖能力的变化、流式细胞术检测凋亡水平的变化；以150 μmol·L^-1吉马酮处理胃癌MGC803、SGC7901细胞48 h，通过Western blotting实验检测HBXIP、PRAS40、p-PRAS40的蛋白表达变化。结果 HBXIP、PRAS40及p-PRAS40在胃癌组织中的表达显著高于癌旁（P＜0.05）并且PRAS40及pPRAS40的表达与HBXIP的表达量成正相关。在胃癌MGC803、SGC7901细胞中沉默HBXIP后，与NC组比较，细胞增殖能力显著降低（P＜0.05） ，细胞凋亡显著增多（P＜0.05） ，PRAS40、p-PRAS40蛋白表达明显降低；而过表达HBXIP后，细胞增殖能力显著增强（P＜0.05） ，PRAS40、p-PRAS40蛋白表达明显升高。与溶剂对照组比较，吉马酮可以剂量相关性地抑制胃癌细胞增殖并促进凋亡（P＜0.05） ，并抑制HBXIP、PRAS40及p-PRAS40的表达。结论 吉马酮通过下调HBXIP促进胃癌细胞增殖，其机制可能与HBXIP下调的PRAS40及其磷酸化蛋白水平有关。

Objective To study the mechanism of the regulation of gastric cancer cell proliferation by germacrone through HBXIP. Methods Gastric cancer tissues and paired paracancer tissues were collected from 20 patients undergoing gastrectomy. The expression of HBXIP, prolin-rich protein kinase B substrate protein (PRAS40) with relative molecular weight of 4×10⁴ and p-PRAS40 were detected by immunohistochemical and Western blotting. The correlation between PRAS40 and p-PRAS40 expression and HBXIP expression in 20 cancer tissues was analyzed by correlation Analyze. After overexpressing or knocking down HBXIP in MGC803 and SGC7901, the change of cell proliferation was detected by MTT. After HBXIP was overexpressed or knocked-down in MGC803 and SGC7901, the apoptosis was detected by flow cytometry and Hoechst 33285, and the expression of HBXIP, PRAS40 and p-PRAS40 were detected by Western blotting. MGC803 and SGC7901 were treated with 0, 50, 100, 150, 200, 250 μmol·L^-1 germacrone for 48 h. The proliferation ability of the cells was detected by MTT and the apoptosis level was detected by flow cytometry. The expression of HBXIP, PRAS40 and p-PRAS40 in MGC803 and SGC7901 were treated with 150 μmol·L^-1 germacrone for 48 h were detected by Western blotting. Results The expression of HBXIP, PRAS40 and p-PRAS40 in gastric cancer tissues was significantly higher than that in adjacent gastric cancer tissues (P＜0.05), and the expression of PRAS40 and p-PRAS40 was positively correlated with the expression level of HBXIP. After knocking down HBXIP in MGC803 and SGC7901, compared to the NC group, cell proliferation was decreased and cell apoptosis was increased (P＜0.05). The expression of PRAS40 and pPRAS40 was also significantly decreased or increased after HBXIP was knocked down or overexpressed in MGC803 and SGC7901. Germacrone inhibited the proliferation and induced apoptosis in a dose-dependent manner (P＜0.05). Germacrone inhibited the expression of HBXIP, PRAS40 and p-PRAS40. Conclusion Germacrone can promote the proliferation of gastric cancer cells by down-regulating HBXIP, and the mechanism may be related to PRAS40 and p-PRAS40 down-regulated by HBXIP.

R285.5

合肥市自然科学基金资助项目（2021012）；安徽省高等学校科学研究项目（自然科学类）（2022AH050712）；合肥市第二人民医院院级课题资助项目（2021ygkt35）