1.大理大学第一附属医院消化内科, 云南 大理 671000; 2.大理州人民医院麻醉科, 云南 大理 671000
R-33
1.Department of Gastroenterology, the First Affiliated Hospital of Dali University, Dali 671000, China. 2. Department of Anesthesiology, Dali people’s Hospital, Dali 671000
目的 研究 miR-572 对胃癌细胞增殖、凋亡的影响和机制。 方法 Real-time PCR 方法测定 miR-572 在胃癌细胞和正常胃黏膜细胞中的表达差异。胃癌细胞 NCI-N87 中转染 miR-572 inhibitor,MTT 测定细胞增殖,PI 单染法测定细胞周期分布,Annexin V-FITC/ PI 法测定细胞凋亡变化,Western blot 检测细胞周期依赖性蛋白激酶 4 (CDK4)、p21、B 细胞淋巴瘤/ 白血病-2(Bcl-2)、Bcl-2 相关 X 蛋白(Bax)蛋白表达。在线靶基因预测软件发现含 WW 域的氧化还原酶(WWOX)可能为 miR-572 的靶基因,荧光素酶报告系统鉴定靶向关系。将 WWOX siRNA 和 miR-572 inhibitor 共转染到胃癌细胞中,利用上述方法测定细胞增殖、周期和凋亡变化。 结果 miR-572 在胃癌细胞中的表达水平明显高于正常胃黏膜细胞。 miR-572 inhibitor 转染后的胃癌细胞增殖能力下降,细胞 G1 期比例升高,细胞凋亡率升高,细胞中 CDK4、Bcl-2 蛋白表达减少,p21、Bax 蛋白表达增多。 miR-572 靶向负调控 WWOX 蛋白表达。 WWOX siRNA 可以逆转 miR-572 inhibitor 对胃癌细胞增殖抑制、周期阻滞和凋亡促进作用。 结论 下调 miR-572 靶向负调控 WWOX 抑制胃癌细胞增殖,阻滞细胞周期并诱导细胞凋亡。
Objective To study the effects and mechanism of microRNA (miR) -572 on the proliferation and apoptosis of gastric cancer cells. Methods Differences in miR-572 expression between gastric cancer cells and normal gastric mucosal cells was detected by Real-time PCR. NCI-N87 cells transfected with the miR-572 inhibitor were examined using the following techniques: MTT assay of cell proliferation, propidium iodide ( PI) single staining to determine cell cycle distribution, annexin V-fluorescein isothiocyanate / PI to measure apoptosis, and Western blot to detect the expression of cyclin-dependent kinase 4 ( CDK4), p21, B-cell lymphoma / leukemia - 2 ( Bcl-2), and Bcl-2-associated X protein (Bax). An online target gene prediction software identified the gene encoding WW domain-containing oxidoreductase (WWOX) as a possible target of miR-572. This relationship was tested using a luciferase reporter system in which WWOX small interfering RNA ( siRNA) and the miR-572 inhibitor were cotransfected into gastric cancer cells, followed by measurement of cell proliferation, cell cycle distribution and apoptosis as above. Results The expression level of miR-572 in gastric cancer cells was significantly higher than that in normal gastric mucosal cells. Gastric cancer cells transfected with the miR-572 inhibitor showed decreased proliferation, an increased proportion of G1 phase, an increased apoptosis rate, decreased expression of CDK4 and Bcl-2, and increased expression of p21 and Bax protein.,m iR-572 negatively regulated the expression of WWOX protein. WWOX siRNA reversed the inhibitory effect of the miR-572 inhibitor on proliferation, cell cycle arrest and apoptosis of gastric cancer cells. Conclusions Downregulation of miR-572-mediated negative regulation of WWOX inhibited the proliferation of gastric cancer cells, blocked the cell cycle and induced apoptosis.
阮庆芬,杨德祥,杨理伟. miR-572 对胃癌细胞增殖和凋亡影响及其作用机制研究[J].中国比较医学杂志,2021,31(3):82~88.
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