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下调MAC30对乳腺癌细胞焦亡、血管生成及ATP1A2蛋白的作用机制
Mechanism of down-regulation of MAC30 on pyrophosis,angiogenesis and ATP1A2 protein in breast cancer cells
- 局解手术学杂志 2023年32卷第6期 页码：481-486
- 作者机构：
1.宣城中心医院心胸乳腺外科，安徽 宣城 242000
2.皖南医学院弋矶山医院甲乳外科，安徽 芜湖 241000
- 作者简介：
陈剑平，E-mail：chenjianping003@163.com
- 基金信息：西藏自治区卫生健康委员会“组援医学项目”专项基金(20202531)
DOI：10.11659/jjssx.09E022120
中图分类号： R737.9纸质出版日期：2023-06-25，
收稿日期：2022-09-22，
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潘蕾蕾,马俊,叶亭亭等.下调MAC30对乳腺癌细胞焦亡、血管生成及ATP1A2蛋白的作用机制[J].局解手术学杂志,2023,32(06):481-486.
PAN Lei-lei,MA Jun,YE Ting-ting,et al.Mechanism of down-regulation of MAC30 on pyrophosis,angiogenesis and ATP1A2 protein in breast cancer cells[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(06):481-486.
潘蕾蕾,马俊,叶亭亭等.下调MAC30对乳腺癌细胞焦亡、血管生成及ATP1A2蛋白的作用机制[J].局解手术学杂志,2023,32(06):481-486. DOI： 10.11659/jjssx.09E022120.
PAN Lei-lei,MA Jun,YE Ting-ting,et al.Mechanism of down-regulation of MAC30 on pyrophosis,angiogenesis and ATP1A2 protein in breast cancer cells[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(06):481-486. DOI： 10.11659/jjssx.09E022120.
摘要
目的
2
探讨下调MAC30对乳腺癌细胞焦亡、血管生成及ATP1A2蛋白水平的影响。
方法
2
MCF-7细胞制成单个细胞悬液，分为乳腺癌组（MCF-7细胞）、NC组（MCF-7细胞转染MAC30-NC-shRNA）、MAC30 shRNA组（MCF-7细胞转染MAC30-shRNA）、ATP1A2组（MCF-7细胞转染ATP1A2-shRNA）及MAC30 shRNA+ATP1A2组（MCF-7细胞转染MAC30-shRNA、ATP1A2-shRNA）。qRT-PCR检测各组MCF-7细胞中MAC30 mRNA表达以证实转染效率，采用CCK-8法检测细胞增殖活性；流式细胞仪检测细胞焦亡率；体外血管形成实验检测血管形成数目；Western blot检测ATP1A2、SCr、AKT及PI3K水平。
结果
2
乳腺癌组与NC组MCF-7细胞中MAC30 mRNA表达水平、MCF-7细胞焦亡率及MCF-7细胞血管形成数目比较，差异均无统计学意义（
P
>
0.05）。与乳腺癌组和NC组比较，MAC30 shRNA组、ATP1A2组及MAC30 shRNA+ATP1A2组MCF-7细胞中MAC30 mRNA表达水平及血管形成数目降低，MCF-7细胞焦亡率升高，差异有统计学意义（
P
<
0.05），其中MAC30 shRNA+ATP1A2组降低/升高最明显（
P
<
0.05）。与NC组相比，MAC30 shRNA组、ATP1A2组及MAC30 shRNA+ATP1A2组细胞增殖活性均降低（
P
<
0.05）。与NC组相比，MAC30 shRNA组ATP1A2、SCr、AKT及PI3K表达水平降低（
P
<
0.05）。
结论
2
下调MAC30可以抑制乳腺癌细胞增殖及血管生成，加快焦亡，其机制与抑制ATP1A2蛋白表达相关。
Abstract
Objective
2
To explore the effect of down-regulation of MAC30 on pyrophosis， angiogenesis and ATP1A2 protein level in breast cancer cells.
Methods
2
MCF-7 cells were made into single cell suspension and divided into the breast cancer group （MCF-7 cells）， the NC group （MCF-7 cells transfected with MAC30-NC-shRNA）， the MAC30 shRNA group （MCF-7 cells transfected with MAC30-shRNA）， the ATP1A2 group （MCF-7 cells transfected with ATP1A2-shRNA） and the MAC30 shRNA +ATP1A2 group （MCF-7 cells transfected with MAC30-shRNA and ATP1A2-shRNA）. qRT-PCR was used to detect the MAC30 mRNA expression in MCF-7 cells of each group to verify the transfection efficiency. CCK-8 assay was used to detect the cell proliferation activity. Flow cytometry was used to detect the pyrophosis rate. In vitro angiogenesis experiment was used to detect the number of angiogenesis. Western blot was used to detect the levels of ATP1A2， SCr， AKT and PI3K.
Results
2
There was no statistically significant difference in the MAC30 mRNA expression level in MCF-7 cells， the pyrophosis rate of MCF-7 cells or the number of angiogenesis in MCF-7 cells between the breast cancer group and the NC group （
P
>
0.05）. Compared with the breast cancer group and the NC group， the MAC30 mRNA expression levels and the number of angiogenesis in MCF-7 cells of the MAC30 shRNA group， the ATP1A2 group， and the MAC30 shRNA+ATP1A2 group were decreased， and the pyrophosis rate in MCF-7 cells was increased， with statistical significance （
P
<
0.05）， of which the MAC30 shRNA+ATP1A2 group decreased/increased most significantly （
P
<
0.05）. Compared with the NC group， the cell proliferative activities of the MAC30 shRNA group， the ATP1A2 group， and the MAC30 shRNA+ATP1A2 group were decreased （
P
<
0.05）. Compared with the NC group， the expression levels of ATP1A2， SCr， AKT， and PI3K of the MAC30 shRNA group were decreased （
P
<
0.05）.
Conclusion
2
Down-regulation of MAC30 can inhibit the proliferation and angiogenesis of breast cancer cells and accelerate pyroptosis， which may be related to the inhibition of ATP1A2 protein expression.
关键词
乳腺癌脑膜瘤相关蛋白焦亡血管生成Na+-K+-ATP酶α2亚基基因
Keywords
breast cancermeningioma-associated proteinpyroptosisangiogenesisNa+-K+-ATPase α2 subunit gene
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