Long non-coding RNA TSI inhibited the metastasis induced by TGF-β1 in breast cencer cell MCF-7 and BT474

doi:10.3969/j.issn.1009-976X.2021.02.008

Abstract

Abstract: Objective To uncover the function of long non-coding RNA TSI （Lnc-TSI） in the TGF-β₁ induced metastasis of breast cancer cell MCF-7 and BT474. Methods We treated the MCF-7 and BT474 cells with TGF-β1 and investigated the morphological changes. Epithelial-mesenchymal transition （EMT） associated proteins were measured with western blot and cell invasion were tested with transwell assay. And then qPCR was used toverify the expression of Lnc-TSI in MCF-10A cells, compared with MCF-7 and BT474 cells with or without TGF-β1 treatment. After knocking out and overexpressing Lnc-TSI in MCF-7 and BT474 cells respectively, we conducted transwell assay to see the changes of cellmigration and invasion. Results Morphology scale showed highly possible EMT changes in MCF-7 and BT474 cells. Also, EMT associated protein E-cadherin were down-regulated while N-cadherin and Vimentin were up-regulated, consisting with the enhanced invasion after treating with TGF-β1. Compared with MCF-10A cells, Lnc-TSI level was much higher in MCF-7 and BT474 cells. TGF-β1 dramatically up-regulated the expression of Lnc-TSI in MCF-7 and BT474 cells. Knocking out and overexpressing Lnc-TSI in MCF-7 and BT474 cells were able to enhanced and inhibited the cell migration and invasion respectively, which were both more obvious with TGF-β1 treatment. Conclusion Lnc-TSI was up-regulated following the metastasis mediated by TGF-β1 in MCF-7 and BT474 cells. Lnc-TSI suppressed the metastasis of MCF-7 and BT474 cells.

Key words: long non-coding RNA, TGF-β1, breast cancer, metastasis

摘要： 目的研究长链非编码RNA TSI（Lnc-TSI）在TGF-β1促进乳腺癌细胞MCF-7和BT474转移中的作用。方法 TGF-β1处理乳腺癌细胞MCF-7和BT474后,观察细胞形态变化,western blot实验检测上皮-间充质转化相关蛋白的变化,transwell实验检测细胞侵袭能力。qRT-PCR分别检测MCF-10A和TGF-β1处理前后MCF-7、BT474细胞的Lnc-TSI变化。在MCF-7和BT474细胞中分别敲除和过表达Lnc-TSI,transwell实验检测细胞在TGF-β1处理后侵袭迁移能力的变化。结果 TGF-β1引起乳腺癌细胞MCF-7和BT474发生上皮-间充质转化,细胞形态呈梭形改变,上皮-间充质转化相关蛋白E-cadherin下调,N-cadherin和Vimentin上调,细胞侵袭数目增加。与正常乳腺上皮细胞MCF-10A细胞相比,MCF-7和BT474细胞中的Lnc-TSI表达更高。TGF-β1处理能显著上调MCF-7和BT474细胞中的Lnc-TSI水平。Transwell实验显示敲除和过表达Lnc-TSI能分别增强和抑制MCF-7和BT474细胞的侵袭和迁移,在TGF-β1处理下这种现象更为明显。结论在TGF-β1促进的乳腺癌细胞MCF-7和BT474的转移进程中Lnc-TSI表达上调,Lnc-TSI能抑制乳腺癌细胞MCF-7和BT474的转移。

关键词: 长链非编码RNA, TGF-β1, 乳腺癌, 转移

CLC Number:

R737.9

TAN Zi-cong, ZHANG Yang-fan, HUANG Xiao-yan, YANG Hao-jie, CAO Ming-hui. Long non-coding RNA TSI inhibited the metastasis induced by TGF-β1 in breast cencer cell MCF-7 and BT474[J]. Lingnan Modern Clinics In Surgery, 2021, 21(02): 171-176.

谭梓聪, 张洋璠, 黄晓燕, 杨浩杰, 曹铭辉. 长链非编码RNA TSI抑制TGF-β1介导的乳腺癌细胞MCF-7和BT474转移[J]. 岭南现代临床外科, 2021, 21(02): 171-176.

References

[1] Siegel RL, Miller KD, Jemal A.Cancer statistics, 2020[J]. CA Cancer J Clin, 2020,70(1):7-30.
[2] Ma Y, Shepherd J, Zhao D, et al.SOX9 Is Essential for Triple-Negative Breast Cancer Cell Survival and Metastasis[J]. Mol Cancer Res, 2020,18(12):1825-1838.
[3] Li J, Huo X, Zhao F, et al.Association of Cyclin-Dependent Kinases 4 and 6 Inhibitors With Survival in Patients With Hormone Receptor-Positive Metastatic Breast Cancer: A Systematic Review and Meta-analysis[J]. JAMA Netw Open, 2020,3(10):e2020312.
[4] Sim WJ, Iyengar PV, Lama D, et al.c-Met activation leads to the establishment of a TGFbeta-receptor regulatory network in bladder cancer progression[J]. Nat Commun, 2019,10(1):4349.
[5] Dongre A, Rashidian M, Eaton EN, et al.Direct and Indirect Regulators of Epithelial-Mesenchymal Transition (EMT)-mediated Immunosuppression in Breast Carcinomas[J]. Cancer Discov, 2021,11(5):1286-1305.
[6] Su S, Liu Q, Chen J, et al.A positive feedback loop between mesenchymal-like cancer cells and macrophages is essential to breast cancer metastasis[J]. Cancer Cell, 2014,25(5):605-620.
[7] Navas T, Kinders RJ, Lawrence SM, et al.Clinical Evolution of Epithelial-Mesenchymal Transition in Human Carcinomas[J]. Cancer Res, 2020,80(2):304-318.
[8] Wang Z, Li Z, Wu Q, et al.DNER promotes epithelial-mesenchymal transition and prevents chemosensitivity through the Wnt/beta-catenin pathway in breast cancer[J]. Cell Death Dis, 2020,11(8):642.
[9] Zhao B, Liu L, Mao J, et al.PIM1 mediates epithelial-mesenchymal transition by targeting Smads and c-Myc in the nucleus and potentiates clear-cell renal-cell carcinoma oncogenesis[J]. Cell Death Dis, 2018,9(3):307.
[10] Wu W, Chen F, Cui X, et al.LncRNA NKILA suppresses TGF-beta-induced epithelial-mesenchymal transition by blocking NF-kappaB signaling in breast cancer[J]. Int J Cancer, 2018,143(9):2213-2224.
[11] Li GY, Wang W, Sun JY, et al.Long non-coding RNAs AC026904.1 and UCA1: a “one-two punch” for TGF-beta-induced SNAI2 activation and epithelial-mesenchymal transition in breast cancer[J]. Theranostics, 2018,8(10):2846-2861.
[12] Mondal T, Subhash S, Vaid R, et al.MEG3 long noncoding RNA regulates the TGF-beta pathway genes through formation of RNA-DNA triplex structures[J]. Nat Commun, 2015,6:7743.
[13] Statello L, Guo CJ, Chen LL, et al.Gene regulation by long non-coding RNAs and its biological functions[J]. Nat Rev Mol Cell Biol, 2021, 22(2): 96-118.
[14] Suwakulsiri W, Chen M, Greening DW, et al.Analysis of Annotated and Unannotated Long Noncoding RNAs from Exosome Subtypes Using Next-Generation RNA Sequencing[J]. Methods Mol Biol, 2021,2254:195-218.
[15] Wang X, Hu X, Song W, et al.Mutual dependency between lncRNA LETN and protein NPM1 in controlling the nucleolar structure and functions sustaining cell proliferation[J]. Cell Res, 2021. doi: 10.1038/s41422-020-00458-6
[16] Hu JJ, Zhou C, Luo X, et al.Linc-SCRG1 accelerates progression of hepatocellular carcinoma as a ceRNA of miR26a to derepress SKP2[J]. J Exp Clin Cancer Res, 2021,40(1):26.
[17] Guo B, Wu S, Zhu X, et al.Micropeptide CIP2A-BP encoded by LINC00665 inhibits triple-negative breast cancer progression[J]. EMBO J, 2020,39(1):e102190.
[18] Wang P, Chen W, Ma T, et al.lncRNA lnc-TSI Inhibits Metastasis of Clear Cell Renal Cell Carcinoma by Suppressing TGF-beta-Induced Epithelial-Mesenchymal Transition[J]. Mol Ther Nucleic Acids, 2020,22:1-16.
[19] Wang P, Luo ML, Song E, et al.Long noncoding RNA lnc-TSI inhibits renal fibrogenesis by negatively regulating the TGF-beta/Smad3 pathway[J]. Sci Transl Med, 2018,10(462).
[20] Xu J, Lamouille S, Derynck R.TGF-beta-induced epithelial to mesenchymal transition[J]. Cell Res, 2009,19(2):156-172.
[21] Hao Y, Baker D, Ten DP.TGF-beta-Mediated Epithelial-Mesenchymal Transition and Cancer Metastasis[J]. Int J Mol Sci, 2019,20(11): 2767.