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罗勇,吴双华,康永清,唐新桥,王泽强,王婷.黄芩苷抑制乳腺癌细胞侵袭、迁移与凋亡的机制研究[J].湖南中医药大学学报,2022,42(1):25-30[点击复制] |
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黄芩苷抑制乳腺癌细胞侵袭、迁移与凋亡的机制研究 |
罗勇,吴双华,康永清,唐新桥,王泽强,王婷 |
(株洲市中心医院, 湖南 株洲 412007) |
摘要: |
目的 用黄芩苷干预人乳腺癌细胞株MCF-7,观察黄芩苷对乳腺癌细胞侵袭、迁移与凋亡的影响及其作用机制。方法 实验分为正常组、模型组、黄芩苷组、miR-126组、LNA组、LNA-126组和LNA-126+黄芩苷组。采用RT-PCR检测miR-126、miR-145、miR-100和let-7c的表达水平,MTT法检测细胞增殖情况,Transwell实验检测细胞侵袭与迁移情况,Western blot法检测p53、Caspase-3、Caspase-9、Bcl-2、Bax和p-p38蛋白的表达水平,流式细胞术检测细胞凋亡情况。结果 RT-PCR结果显示,与模型组相比,黄芩苷作用于MCF-7细胞后miR-126、miR-145、miR-100和let-7c表达明显上调(P<0.05,P<0.01,P<0.001),其中以miR-126最为显著(P<0.001);与正常组相比,4种miRNA在模型组中的表达均下调,其中以miR-126下调最为明显(P<0.05);MTT结果显示,MCF-7细胞的增殖受到miR-126和黄芩苷的抑制(P<0.05),而MCF-7细胞的增殖受到LNA-126的促进(P<0.05);Transwell实验结果显示,MCF-7细胞的侵袭和迁移受到miR-126和黄芩苷的抑制(P<0.05),而黄芩苷对MCF-7细胞侵袭和迁移的抑制受到LNA-126的拮抗(P>0.05);Western blot结果显示,黄芩苷及miR-126作用于MCF-7细胞后Bcl-2表达水平下降(P<0.05),p53、Caspase-3、Caspase-9、Bax及p-p38表达水平上升(P<0.05);流式细胞术结果显示,黄芩苷及miR-126促进MCF-7细胞凋亡(P<0.05)。结论 黄芩苷可以抑制乳腺癌细胞的增殖、侵袭与迁移,并促进其凋亡,其机制可能与通过上调miR-126有关。 |
关键词: 乳腺癌 黄芩苷 miR-126 侵袭与迁移 细胞凋亡 |
DOI:10.3969/j.issn.1674-070X.2022.01.006 |
投稿时间:2021-07-28 |
基金项目:湖南省自然科学基金项目(2018JJ4096)。 |
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Mechanism of baicalin inhibiting invasion, migration and apoptosis of breast cancer cells |
LUO Yong,WU Shuanghua,KANG Yongqing,TANG Xinqiao,WANG Zeqiang,WANG Ting |
(Central Hospital of Zhuzhou, Zhuzhou, Hunan 412007, China) |
Abstract: |
Objective Human breast cancer cell line MCF-7 was treated with baicalin to observe the effect and mechanism of baicalin on invasion, migration and apoptosis of breast cancer cells. Methods The experiment was divided into control group, model group, baicalin group, miR-126 group, LNA group, LNA-126 group and LNA-126+baicalin group. The expression levels of miR-126, miR-145, miR-100, let-7c were detected by RT-PCR, and the cell proliferation was determined by using MTT method. The invasion and migration of cells were detected by Transwell test, and the expression levels of p53, Caspase-3, Caspase-9, Bcl-2, Bax and p-p38 were determined by Western blot. Cell apoptosis was detected by flow cytometry. Results The results of RT-PCR showed that compared with model group, the expression levels of miR-126, miR-145, miR-100, and let-7c were significantly up-regulated after baicalin was treated on MCF-7 cells (P<0.05, P<0.01, P<0.001), especially miR-126 (P<0.001); compared with control group, the expression levels of four miRNAs in model group were down regulated, especially miR-126 (P<0.05); MTT results showed that miR-126 and baicalin can inhibit the growth of MCF-7 cells (P<0.05), while LNA-126 may promote the growth MCF-7 cells (P<0.05). Transwell test results showed that miR-126 and baicalin can inhibit the invasion and migration of MCF-7 cells (P<0.05), while LNA-126 can antagonize the inhibitory effect of baicalin (P>0.05). Western blot results showed that the expression level of Bcl-2 decreased after baicalin and miR-126 acted on MCF-7 cells (P<0.05), and the expression levels of p53, Caspase-3, Caspase-9, Bax, and p-p38 increased (P<0.05). The results of flow cytometry showed that baicalin and miR-126 promoted the apoptosis of MCF-7 cells (P<0.05). Conclusion Baicalin can inhibit the proliferation, invasion and migration of breast cancer cells, and promote their apoptosis. The mechanism may be related to the up-regulation of miR-126. |
Key words: breast cancer baicalin miR-126 invasion and migration apoptosis |
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