[1]朱方园,邵营波,陈　琦,等.长链非编码RNA前列腺癌相关转录本7对三阴性乳腺癌MDA-MB-436细胞增殖、迁移及侵袭的影响[J].新乡医学院学报,2022,39(8):701-707.[doi:10.7683/xxyxyxb.2022.08.001]
　ZHU Fangyuan,SHAO Yingbo,CHEN Qi,et al.Effect of long non-coding RNA prostate cancer associated transcript 7 on proliferation,migration and invasion of triple-negative breast cancer MDA-MB-436 cells[J].Journal of Xinxiang Medical University,2022,39(8):701-707.[doi:10.7683/xxyxyxb.2022.08.001]
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长链非编码RNA前列腺癌相关转录本7对三阴性乳腺癌MDA-MB-436细胞增殖、迁移及侵袭的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
39
期数:
2022年8
页码:
701-707
栏目:
基础研究
出版日期:
2022-08-05

文章信息/Info

Title:
Effect of long non-coding RNA prostate cancer associated transcript 7 on proliferation,migration and invasion of triple-negative breast cancer MDA-MB-436 cells
作者:
朱方园; 邵营波; 陈　琦; 贺亚宁; 刘朝俊; 聂　冰; 刘　慧
（郑州大学人民医院/河南省人民医院乳腺外科，河南　郑州　450003）
Author(s):
ZHU Fangyuan; SHAO Yingbo; CHEN Qi; HE Yaning; LIU Chaojun; NIE Bing; LIU Hui
(Department of Breast Surgery,People′s Hospital of Zhengzhou University/Henan Provincial People′s Hospital,Zhengzhou 450003,Henan Province,China)
关键词:
长链非编码RNA; 前列腺癌相关转录本7; 三阴性乳腺癌; 细胞增殖; 细胞周期
Keywords:
long non-coding RNA; prostate cancer associated transcript 7; triple-negative breast cancer; cell proliferation; migration; invasion; cell cycle
分类号:
R737.9
DOI:
10.7683/xxyxyxb.2022.08.001
文献标志码:
A
摘要:
目的　探讨长链非编码RNA前列腺癌相关转录本7（lncRNA PCAT7）对三阴性乳腺癌MDA-MB-436细胞增殖、迁移及侵袭的影响。方法　设计合成lncRNA PCAT7-siRNA1、lncRNA PCAT7-siRNA2、lncRNA PCAT7-siRNA3及control siRNA。MDA-MB-436细胞培养至细胞融合度达80%时，将细胞分为lncRNA PCAT7-siRNA1组、lncRNA PCAT7-siRNA2组、lncRNA PCAT7-siRNA3组3个干扰组和1个空载组，分别转染lncRNA PCAT7-siRNA1、lncRNA PCAT7-siRNA2、lncRNA PCAT7-siRNA3和control siRNA，转染6 h后细胞用完全培养基继续培养48 h，收集各组细胞，采用实时聚合酶链反应法检测转染后各组细胞lncRNA PCAT7表达情况，选择lncRNA PCAT7表达水平最低的lncRNA PCAT7-siRNA3组MDA-MB-436细胞用于后续实验。采用克隆形成实验检测lncRNA PCAT7-siRNA3组和空载组MDA-MB-436细胞的增殖能力，划痕实验检测lncRNA PCAT7-siRNA3组和空载组MDA-MB-436细胞的迁移能力，Transwell小室实验检测lncRNA PCAT7-siRNA3组和空载组MDA-MB-436细胞的侵袭能力，流式细胞术检测lncRNA PCAT7-siRNA3组和空载组MDA-MB-436细胞周期分布情况，Western blot法检测lncRNA PCAT7-siRNA3组和空载组MDA-MB-436细胞p27蛋白表达情况。结果　lncRNA PCATT-siRNA1干扰组MDA-MB-436细胞中lncRNA PCAT7 mRNA相对表达量与空载组比较差异无统计学意义（t=-4.286,P>0.05），lncRNA PCAT7-siRNA2组、lncRNA PCAT7-siRNA3组MDA-MB-436细胞中lncRNA PCAT7 mRNA相对表达量显著低于空载组（t=-28.028、-47.748,P<0.05），lncRNA PCAT7-siRNA3组MDA-MB-436细胞中lncRNA PCAT7 mRNA相对表达量显著低于lncRNA PCAT7-siRNA2组（t=12.705,P<0.01），选用转染lncRNA PCAT7-siRNA3组MDA-MB-436细胞进行后续实验。培养14 d后，lncRNA PCAT7-siRNA3组MDA-MB-436细胞克隆形成数显著少于空载组（t=-7.434,P<0.01）。细胞划痕培养48 h后，lncRNA PCAT7-siRNA3组MDA-MB-436细胞划痕愈合率显著低于空载组（t=-11.340,P<0.01）。lncRNA PCAT7-siRNA3组MDA-MB-436细胞侵袭数显著少于空载组（t=-7.120，P<0.01）。lncRNA PCAT7-siRNA3组MDA-MB-436组细胞G0/G1期细胞比例显著高于空载组，G2/M期细胞比例显著低于空载组（t=4.920、-6.990,P<0.01），lncRNA PCAT7-siRNA3组与空载组MDA-MB-436细胞S期细胞比例比较差异无统计学意义（t=0.083,P>0.05）。lncRNA PCAT7-siRNA3组MDA-MB-436细胞中p27蛋白相对表达量显著高于空载组（t=3.200，P<0.05）。结论　下调lncRNA PCAT7可抑制三阴性乳腺癌MDA-MB-436细胞的增殖、迁移与侵袭能力，其调控机制可能与增加细胞周期抑制蛋白p27表达及阻滞细胞周期有关。
Abstract:
Objective　To investigate the effect of long non-coding RNA prostate cancer associated transcript 7 (lncRNA PCAT7) on the proliferation,migration and invasion of triple-negative breast cancer MDA-MB-436 cells.Methods　The lncRNA PCAT7-siRNA1,lncRNA PCAT7-siRNA2,lncRNA PCAT7-siRNA3 and control siRNA were designed and synthesized.When MDA-MB-436 cells were cultured until the degree of cell fusion reached 80%,the cells were divided into lncRNA PCAT7-siRNA1 group,lncRNA PCAT7-siRNA2 group,lncRNA PCAT7-siRNA3 group and empty group,and lncRNA PCAT7-siRNA1,lncRNA PCAT7-siRNA2,lncRNA PCAT7-siRNA3 and control siRNA were transfected,respectively.At six hours after transfection,the cells were continually cultured for 48 hours with complete medium.The expression of lncRNA PCAT7 in cells in the each group was detected by real-time polymerase chain reaction.MDA-MB-436 cells with the lowest lncRNA PCAT7 expression in the interference group were selected for subsequent experiment.The proliferation ability of MDA-MB-436 cells in the lncRNA PCAT7-siRNA interference group and the empty group was detected by colony formation assay.The migration ability of MDA-MB-436 cells in the lncRNA PCAT7-siRNA interference group and the empty group was detected by cell scratch assay.The invasion ability of MDA-MB-436 cells in the lncRNA PCAT7-siRNA interference group and the empty group was detected by transwell chamber assay.The cell cycle distribution of MDA-MB-436 cells in the lncRNA PCAT7-siRNA interference group and the empty group was detected by flow cytometry.The relative expression of p27 protein in MDA-MB-436 cells in the lncRNA PCAT7-siRNA interference group and the empty group was determined by Western blot.Results　There was no significant difference in the relative expression of lncRNA PCAT7 mRNA in MDA-MB-436 cells between the lncRNA PCAT7-siRNA1 group and the empty group (t=-4.286,P>0.05),the relative expression of lncRNA PCAT7 in the lncRNA PCAT7-siRNA2 group and lncRNA PCAT7-siRNA3 group was significantly lower than that in the empty group (t=-28.028,-47.748;P<0.05),and the relative expression of lncRNA PCAT7 in the lncRNA PCAT7-siRNA3 group was significantly lower than that in the lncRNA PCAT7-siRNA2 group (t=12.705,P<0.01).Therefore,MDA-MB-436 cells transfected with lncRNA PCAT7-siRNA3 were selected for subsequent experiments.After 14 days of culture,the number of colon of MDA-MB-436 cells in the lncRNA PCAT7-siRNA3 group was significantly lower than that in the empty group (t=-7.434,P<0.01).After 48 hours of culture in cell scratch assay,the scratch healing rate of MDA-MB-436 cells in the lncRNA PCAT7-siRNA3 group was significantly lower than that in the empty group (t=-11.340,P<0.01).The number of MDA-MB-436 cells invasion in the lncRNA PCAT7-siRNA3 group was significantly lower than that in the empty group (t=-7.120,P<0.01).The proportion of MDA-MB-436 cells at G0/G1 phase in the lncRNA PCAT7-siRNA3 group was significantly higher than that in the empty group,and the proportion of MDA-MB-436 cells at G2/M phase was significantly lower than that in the empty group (t=4.920,-6.990;P<0.01).There was no significant difference in the proportion of MDA-MB-436 cells at S phase between the lncRNA PCAT7-siRNA3 group and the empty group (t=0.083,P>0.05).The relative expression level of p27 protein in the lncRNA PCAT7-siRNA3 group was significantly higher than that in the empty group (t=3.200,P<0.05).Conclusion　　Down-regulation of lncRNA PCAT7 can inhibit the proliferation,migration and invasion of triple-negative breast cancer MDA-MB-436 cells,and its regulatory mechanisms may be related to the up-regulation of cell cycle inhibitory protein p27 and the cell cycle arrest.

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更新日期/Last Update: 2022-08-05