Effects of TROP-2 gene on adhesion and invasion of human breast cancer cells
Yumin Li, Hongfeng Zhan, Dandan Gong, Zhiyuan Qiu, Yongjing Zhou, Xiudi Xiao, Zhengyan Wu, Yu Fan
Abstract
Yumin Li, Hongfeng Zhan, Dandan Gong, Zhiyuan Qiu, Yongjing Zhou, Xiudi Xiao, Zhengyan Wu, Yu Fan
Abstract
Objective To study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA (siRNA) on adhesion and invasion of human breast cancer cells.Methods Real time PCR was used to detect the TROP-2 mRNA of seven human breast cancer cell lines:Bcap-37,LCC1,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,and ZR75-1.The cell line with hifhest TROP-2 expression was transfected with different doses of TROP-2 siRNA.The expression of TROP-2 mRNA and protein was detected by real-time quantitative polymerase chain reaction (PCR) and immumoflurescence method.The cell adhesion was evaluated by methyl thiazol tetrazolium (MTT) assay,and invasion was exmined by boyden chamber,respectively.Results The TROP-2 mRNA in Bcap-37,LCC1,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468 and ZR75-1 cell lines was 1.362 ±0.057,2.207 ± 0.056,2.997 ± 0.052,0.136 ± 0.045,0.122 ± 0.025,0.194 ± 0.028 and 2.706 ± 0.039 respectively,and MCF-7 showed the highest elevation of TROP-2 mRNA.The real-time quantitative PCR and immumoflurescence method revealed that the expression of TROP-2 mRNA and protein was reduced in a time- and dose-dependent manner (P < 0.01 ;P < 0.01 ).The adhesive rate in siRNA groups (5,10 and 20 nmol/L) was (52.9±2.5)%,(25.6±2.3)% and (12.8±2.2)% (P<0.01),respectively.The transwell results showed that the invasion cells were 78 ± 17,39 ± 15,19 ± 16,136 ± 25 and 139 ± 21 in different groups (5,10,20 umol/L siRNA,and controls),respectively (P <0.01 ).Conclusion TROP-2 gene might play an important role in adhesion and invasion of human breast cancer cells.siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cells. Key words: Breast carcinoma; Tumor-associated calcium signal transducer-2; RNA interference; Invasion
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Objective To study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA (siRNA) on adhesion and invasion of human breast cancer cells.Methods Real time PCR was used to detect the TROP-2 mRNA of seven human breast cancer cell lines:Bcap-37,LCC1,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,and ZR75-1.The cell line with hifhest TROP-2 expression was transfected with different doses of TROP-2 siRNA.The expression of TROP-2 mRNA and protein was detected by real-time quantitative polymerase chain reaction (PCR) and immumoflurescence method.The cell adhesion was evaluated by methyl thiazol tetrazolium (MTT) assay,and invasion was exmined by boyden chamber,respectively.Results The TROP-2 mRNA in Bcap-37,LCC1,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468 and ZR75-1 cell lines was 1.362 ±0.057,2.207 ± 0.056,2.997 ± 0.052,0.136 ± 0.045,0.122 ± 0.025,0.194 ± 0.028 and 2.706 ± 0.039 respectively,and MCF-7 showed the highest elevation of TROP-2 mRNA.The real-time quantitative PCR and immumoflurescence method revealed that the expression of TROP-2 mRNA and protein was reduced in a time- and dose-dependent manner (P < 0.01 ;P < 0.01 ).The adhesive rate in siRNA groups (5,10 and 20 nmol/L) was (52.9±2.5)%,(25.6±2.3)% and (12.8±2.2)% (P<0.01),respectively.The transwell results showed that the invasion cells were 78 ± 17,39 ± 15,19 ± 16,136 ± 25 and 139 ± 21 in different groups (5,10,20 umol/L siRNA,and controls),respectively (P <0.01 ).Conclusion TROP-2 gene might play an important role in adhesion and invasion of human breast cancer cells.siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cells. Key words: Breast carcinoma; Tumor-associated calcium signal transducer-2; RNA interference; Invasion
Key concepts: Molecular biology, Transfection, Cell culture, Messenger RNA, Small interfering RNA, MTT assay, Real-time polymerase chain reaction, Cancer cell