目的 探討氧化苦參堿通過調(diào)節(jié)miR-155-5p表達對胃癌細胞的增殖、凋亡、遷移、侵襲中的作用和其潛在機制。方法 使用TRIzol法提取組織標本和細胞系的總RNA，檢測胃癌組織與癌旁組織中的miR-155-5p表達情況。采用qRT-PCR評估m(xù)iR-155-5p在人胃癌細胞系（SGC7901、MGC803）與正常胃黏膜上皮（GES-1、AGS）細胞中的表達情況。將MGC803分為空白組、轉染錯義序列siRNA組、miR-155-5p模擬物組、miR-155-5p抑制劑組、氧化苦參堿（100 μmol/L）組、氧化苦參堿＋miR-155-5p模擬物組。采用MTT法觀察不同干預下胃癌細胞的生長抑制作用，用細胞集落形成試驗檢測不同干預下胃癌細胞的增殖情況。用流式細胞技術分析不同干預對胃癌細胞凋亡的影響。Transwell法檢測不同干預后胃癌細胞的遷移、侵襲情況。蛋白質(zhì)印跡法檢測MGC803細胞中Claudin 1、c-Myc、cyclin D1、Bcl-2、Caspase-3蛋白的相對表達量。結果 與癌旁組織和胃黏膜細胞（GES-1、AGS）相比，在胃癌組織和胃癌細胞系（SGC7901、MGC803）中miR-155-5p相對表達量升高（P＜0.001）。而氧化苦參堿通過調(diào)節(jié)miR-155-5p表達抑制胃癌細胞生長。MTT實驗表明，miR-155-5p模擬物的轉染后MGC803細胞活力顯著增加，而氧化苦參堿可顯著抑制胃癌細胞、轉染和miR-155-5p模擬物的細胞活力（P＜0.001）。與空白組、錯義序列siRNA組相比，miR-155-5p模擬物轉染后MGC803細胞活力顯著增加，細胞集落生成增加，而氧化苦參堿可顯著抑制胃癌細胞的活力和轉染miR-155-5p模擬物的細胞活力，及細胞集落生成數(shù)（P＜0.001）。miR-155-5p-模擬物組的細胞遷移和侵襲細胞數(shù)顯著增加、凋亡比例降低，而氧化苦參堿組和miR-155-5p-抑制劑組細胞遷移和侵襲細胞數(shù)量降低、凋亡比例升高（P＜0.001）。miR-155-5p模擬組的Claudin 1、c-Myc、Cyclin D1、Bcl-2相對表達量增加，而Caspase-3相對表達量降低（P＜0.01、0.001）。而miR-155-5p抑制劑組、氧化苦參堿組和氧化苦參堿＋miR-155-5p抑制劑組的Claudin 1、c-Myc、Cyclin D1、Bcl-2表達較miR-155-5p模擬組顯著下降，Caspase-3表達增加（P＜0.001）。結論 miR-155-5p可能是胃癌的治療靶點，氧化苦參堿可通過miR-155-5p的調(diào)節(jié)作用發(fā)揮抗腫瘤作用。

Objective To investigate the role and potential mechanism of oxymatrine in regulating the expression of miR-155-5p in the proliferation, apoptosis, migration, and invasion of gastric cancer cells. Method Total RNA from tissue specimens and cell lines was extracted by TRIzol method, to detect the expression of miR-155-5p in gastric cancer tissues and adjacent tissues. The expression of miR-155-5p in human gastric cancer cell lines (SGC7901, MGC803) and normal gastric mucosal epithelial cells (GES-1, AGS) was evaluated by qRT-PCR. MGC803 was divided into blank group, transfected missense sequence siRNA group, miR-155-5p simulant group, miR-155-5p inhibitor group, oxymatrine (100 μmol/L) group, and oxymatrine + miR-155-5p mimics group. MTT assay was used to observe the growth inhibition of gastric cancer cells under different interventions, and colony formation assay was used to detect the proliferation of gastric cancer cells under different interventions. Effects of different interventions on apoptosis of gastric cancer cells were analyzed by flow cytometry. The migration and invasion of gastric cancer cells after different interventions were detected by Transwell method. The relative expression levels of Claudin 1, c-Myc, Cyclin D1, Bcl-2, and Caspase-3 in MGC803 cells were detected by Western blotting. Results The relative expression of miR-155-5p was increased in gastric cancer tissues and gastric cancer cell lines (SGC7901, MGC803) compared with para-cancerous tissues and gastric mucosa cells (GES-1, AGS) (P < 0.001). Oxymatrine inhibited the growth of gastric cancer cells by regulating the expression of miR-155-5p. MTT assay showed that MGC803 cell viability was significantly increased after transfection with miR-155-5p mimicry, and the cell viability of gastric cancer cells, transfection and miR-155-5p mimicry was significantly inhibited by oxymatrine (P < 0.001). Compared with blank group and missense sequence siRNA group, MGC803 cell viability and colony generation were significantly increased after transfection with miR-155-5p mimics, while oxymatrine significantly inhibited the viability of gastric cancer cells, the cell viability and colony generation number of transfected miR-155-5p mimics (P < 0.001). The number of cell migration and invasion cells increased significantly and the proportion of apoptosis decreased in the miR-155-5p simulant group, while the number of cell migration and invasion cells decreased and the proportion of apoptosis increased in the oxymatrine and miR-155-5p- inhibitor groups (P < 0.001). The relative expressions of Claudin 1, c-Myc, Cyclin D1, and Bcl-2 in miR-155-5p simulant group were increased, while the relative expressions of Caspase-3 were decreased (P < 0.01, 0.001). The expressions of Claudin 1, c-Myc, Cyclin D1, and Bcl-2 in miR-155-5p inhibitor group, oxymatrine group, and oxymatrine + miR-155-5p inhibitor group were significantly decreased compared with those in miR-155-5p simulant group, and the expression of Caspase-3 was increased (P < 0.001). Conclusion MiR-155-5p may be a therapeutic target for gastric cancer, and oxymatrine can exert anti-tumor effects through the regulatory effect of miR-155-5p.

R966；R979.1

三二〇一醫(yī)院科研基金資助項目（3201yk201530）