目的 研究吉馬酮通過HBXIP調(diào)控PRAS40/p-PRAS4影響胃癌細胞增殖的機制。方法 收集20例行胃癌切除術(shù)患者的胃癌組織及配對的癌旁組織，通過免疫組化實驗及Western blotting實驗檢測乙型肝炎X相互作用蛋白（HBXIP）、相對分子質(zhì)量為4×104的富含脯氨酸蛋白激酶B底物蛋白（PRAS40）、磷酸化PRAS40（p-PRAS40）的表達量；以20例癌組織中HBXIP、PRAS40、p-PRAS40蛋白表達的免疫組織化學染色評分為數(shù)據(jù)，通過Correlation matrix分析PRAS40、p-PRAS40的表達與HBXIP表達的相關(guān)性；在胃癌MGC803、SGC7901細胞中通過轉(zhuǎn)染質(zhì)粒過表達或沉默HBXIP后（NC組轉(zhuǎn)染空白質(zhì)粒），MTT實驗檢測細胞增殖能力的變化，通過流式細胞術(shù)及Hoechst 33285染色檢測細胞凋亡情況，并通過Westernblotting實驗檢測HBXIP、PRAS40、p-PRAS40的蛋白表達量；以0（溶劑對照組）、50、100、150、200、250 μmol·L^-1吉馬酮處理胃癌MGC803、SGC7901細胞48 h，MTT實驗檢測細胞增殖能力的變化、流式細胞術(shù)檢測凋亡水平的變化；以150 μmol·L^-1吉馬酮處理胃癌MGC803、SGC7901細胞48 h，通過Western blotting實驗檢測HBXIP、PRAS40、p-PRAS40的蛋白表達變化。結(jié)果 HBXIP、PRAS40及p-PRAS40在胃癌組織中的表達顯著高于癌旁（P＜0.05）并且PRAS40及pPRAS40的表達與HBXIP的表達量成正相關(guān)。在胃癌MGC803、SGC7901細胞中沉默HBXIP后，與NC組比較，細胞增殖能力顯著降低（P＜0.05） ，細胞凋亡顯著增多（P＜0.05） ，PRAS40、p-PRAS40蛋白表達明顯降低；而過表達HBXIP后，細胞增殖能力顯著增強（P＜0.05） ，PRAS40、p-PRAS40蛋白表達明顯升高。與溶劑對照組比較，吉馬酮可以劑量相關(guān)性地抑制胃癌細胞增殖并促進凋亡（P＜0.05） ，并抑制HBXIP、PRAS40及p-PRAS40的表達。結(jié)論 吉馬酮通過下調(diào)HBXIP促進胃癌細胞增殖，其機制可能與HBXIP下調(diào)的PRAS40及其磷酸化蛋白水平有關(guān)。

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

合肥市自然科學基金資助項目（2021012）；安徽省高等學?？茖W研究項目（自然科學類）（2022AH050712）；合肥市第二人民醫(yī)院院級課題資助項目（2021ygkt35）