Inhibition of miR‑96 enhances the sensitivity of colorectal cancer cells to oxaliplatin by targeting TPM1

Ge,Tingrui; Xiang,Ping; Mao,Haibing; Tang,Shumin; Zhou,Jinyi; Zhang,Yonggang

doi:10.3892/etm.2020.8936

Inhibition of miR‑96 enhances the sensitivity of colorectal cancer cells to oxaliplatin by targeting TPM1

Authors:
- Tingrui Ge
- Ping Xiang
- Haibing Mao
- Shumin Tang
- Jinyi Zhou
- Yonggang Zhang
View Affiliations

Affiliations: Department of Colorectal Surgery, The First People's Hospital of Lianyungang, Lianyungang, Jiangsu 222002, P.R. China
Published online on: June 25, 2020 https://doi.org/10.3892/etm.2020.8936
Pages: 2134-2140
Copyright: © Ge et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Colorectal cancer (CRC) is one of the major threats to human health worldwide. In the treatment of CRC, chemoresistance affects the efficacy of platinum‑based therapies. Oxaliplatin is one of the most commonly used first‑line medications for the treatment of CRC; however, chemoresistance is common among patients receiving oxaliplatin treatment, which significantly decreases its therapeutic efficacy. The present study focused on the roles of microRNA (miR)‑96 in the oxaliplatin resistance of CRC cells and the underlying mechanisms. First, the expression of miR‑96 was compared between CRC and adjacent tissues. Furthermore, target genes of miR‑96 were predicted, and a dual‑luciferase reporter assay was employed to confirm whether the candidate tropomyosin 1 (TPM1) is a direct target of miR‑96. In addition, CRC cells were transfected with miR‑96 inhibitor, miR‑negative control, small interfering RNA (siRNA) targeting TPM1 or siRNA NC, and then treated with oxaliplatin. CCK‑8 assay and flow cytometry were performed to examine the proliferation and apoptosis of the CRC cell line SW480. Next, reverse transcription‑quantitative PCR and western blot analysis were performed to determine the mRNA and/or protein levels of miR‑96, Bcl‑2, BAX and TPM1. The results indicated that miR‑96 was upregulated in CRC compared with normal adjacent tissues, while TPM1 was downregulated. The luciferase activity was reduced following transfection with miR‑96 mimics and luciferase reporter plasmid containing the wild‑type sequence of the 3'‑untranslated region of TPM1. Furthermore, knockdown of miR‑96 combined with oxaliplatin reduced the viability and induced apoptosis of CRC cells, which was further verified by decreased expression of Bcl‑2 and the increased expression of TPM1 and BAX. Taken together, the downregulation of miR‑96 enhanced the sensitivity of CRC cells to oxaliplatin.

View Figures

View References

1	Liebs S, Keilholz U, Kehler I, Schweiger C, Hayback J and Nonnenmacher A: Detection of mutations in circulating cell-free DNA in relation to disease stage in colorectal cancer. Cancer Med. 8:3761–3769. 2019.PubMed/NCBI View Article : Google Scholar
2	Lei L, Zhao X, Liu S, Cao Q, Yan B and Yang J: MicroRNA-3607 inhibits the tumorigenesis of colorectal cancer by targeting DDI2 and regulating the DNA damage repair pathway. Apoptosis. 24:662–672. 2019.PubMed/NCBI View Article : Google Scholar
3	Wang Q, Wei J, Wang C, Zhang T, Huang D, Wei F, He F, Cai W, Yang P, Zeng S, et al: Gambogic acid reverses oxaliplatin resistance in colorectal cancer by increasing intracellular platinum levels. Oncol Lett. 16:2366–2372. 2018.PubMed/NCBI View Article : Google Scholar
4	Chibaudel B, Tournigand C, Bonnetain F, Maindrault-Goebel F, Lledo G, André T, Larsen AK, Bengrine-Lefevre L, Louvet C and de Gramont A: Platinum-sensitivity in metastatic colorectal cancer: Towards a definition. Eur J Cancer. 49:3813–3820. 2013.PubMed/NCBI View Article : Google Scholar
5	Huang CY, Chiang SF, Chen WT, Ke TW, Chen TW, You YS, Lin CY, Chao KSC and Huang CY: HMGB1 promotes ERK-mediated mitochondrial Drp1 phosphorylation for chemoresistance through RAGE in colorectal cancer. Cell Death Dis. 9(1004)2018.PubMed/NCBI View Article : Google Scholar
6	Ren J, Ding L, Zhang D, Shi G, Xu Q, Shen S, Wang Y, Wang T and Hou Y: Carcinoma-associated fibroblasts promote the stemness and chemoresistance of colorectal cancer by transferring exosomal lncRNA H19. Theranostics. 8:3932–3948. 2018.PubMed/NCBI View Article : Google Scholar
7	Zhang Z, Feng L, Liu P and Duan W: ANRIL promotes chemoresistance via disturbing expression of ABCC1 by regulating the expression of Let-7a in colorectal cancer. Biosci Rep. 38(pii: BSR20180620)2018.PubMed/NCBI View Article : Google Scholar
8	Gupta P, Sata TN, Yadav AK, Mishra A, Vats N, Hossain MM, Sanal MG and Venugopal SK: TGF-β induces liver fibrosis via miRNA-181a-mediated down regulation of augmenter of liver regeneration in hepatic stellate cells. PLoS One. 14(e0214534)2019.PubMed/NCBI View Article : Google Scholar
9	Huerta-Zavala ML, Lopez-Castillejos ES, Requenez-Contreras JL, Granados-Riveron JT and Aquino-Jarquin G: A single miRNA and miRNA sponge expression system for efficient modulation of miR-223 availability in mammalian cells. J Gene Med. 21(e3100)2019.PubMed/NCBI View Article : Google Scholar
10	Wu T, Cao Y, Yang Y and Zhang X, Wang S, Xu LP and Zhang X: A three-dimensional DNA walking machine for the ultrasensitive dual-modal detection of miRNA using a fluorometer and personal glucose meter. Nanoscale. 11:11279–11284. 2019.PubMed/NCBI View Article : Google Scholar
11	Powrozek T, Brzozowska A, Mazurek M, Mlak R, Sobieszek G and Malecka-Massalska T: Combined analysis of miRNA-181a with phase angle derived from bioelectrical impedance predicts radiotherapy-induced changes in body composition and survival of male patients with head and neck cancer. Head Neck. 41:3247–3257. 2019.PubMed/NCBI View Article : Google Scholar
12	Vos PD, Leedman PJ, Filipovska A and Rackham O: Modulation of miRNA function by natural and synthetic RNA-binding proteins in cancer. Cell Mol Life Sci. 76:3745–3752. 2019.PubMed/NCBI View Article : Google Scholar
13	Xie W, Sun F, Chen L and Cao X: miR-96 promotes breast cancer metastasis by suppressing MTSS1. Oncol Lett. 15:3464–3471. 2018.PubMed/NCBI View Article : Google Scholar
14	Bao YH, Wang Y, Liu Y, Wang S and Wu B: MiR-96 expression in prostate cancer and its effect on the target gene regulation. Eur Rev Med Pharmacol Sci. 21:4548–4556. 2017.PubMed/NCBI
15	Hong Y, Liang H, Uzair Ur Rehman, Wang Y, Zhang W, Zhou Y, Chen S, Yu M, Cui S, Liu M, et al: miR-96 promotes cell proliferation, migration and invasion by targeting PTPN9 in breast cancer. Sci Rep. 6(37421)2016.PubMed/NCBI View Article : Google Scholar
16	Wu L, Pu X, Wang Q, Cao J, Xu F, Xu LI and Li K: miR-96 induces cisplatin chemoresistance in non-small cell lung cancer cells by downregulating SAMD9. Oncol Lett. 11:945–952. 2016.PubMed/NCBI View Article : Google Scholar
17	Rapti SM, Kontos CK, Papadopoulos IN and Scorilas A: High miR-96 levels in colorectal adenocarcinoma predict poor prognosis, particularly in patients without distant metastasis at the time of initial diagnosis. Tumour Biol. 37:11815–11824. 2016.PubMed/NCBI View Article : Google Scholar
18	Ress AL, Stiegelbauer V, Winter E, Schwarzenbacher D, Kiesslich T, Lax S, Jahn S, Deutsch A, Bauernhofer T, Ling H, et al: MiR-96-5p influences cellular growth and is associated with poor survival in colorectal cancer patients. Mol Carcinog. 54:1442–1450. 2015.PubMed/NCBI View Article : Google Scholar
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
20	Wu J, Wang F, Liu X, Zhang T, Liu F, Ge X, Mao Y and Hua D: Correlation of IDH1 and B7H3 expression with prognosis of CRC patients. Eur J Surg Oncol. 44:1254–1260. 2018.PubMed/NCBI View Article : Google Scholar
21	Fang L, Li H, Wang L, Hu J, Jin T, Wang J and Yang BB: MicroRNA-17-5p promotes chemotherapeutic drug resistance and tumour metastasis of colorectal cancer by repressing PTEN expression. Oncotarget. 5:2974–2987. 2014.PubMed/NCBI View Article : Google Scholar
22	Zhang JR, Zhu RH and Han XP: MiR-410 affects the proliferation and apoptosis of lung cancer A549 cells through regulation of SOCS3/JAK-STAT signaling pathway. Eur Rev Med Pharmacol Sci. 22:5994–6001. 2018.PubMed/NCBI View Article : Google Scholar
23	Jiang QQ and Liu WB: miR-25 promotes melanoma progression by regulating RNA binding motif protein 47. Med Sci (Paris). 34 Focus issue F1:59–65. 2018.PubMed/NCBI View Article : Google Scholar
24	Guo Y, Pang Y, Gao X, Zhao M, Zhang X, Zhang H, Xuan B and Wang Y: MicroRNA-137 chemosensitizes colon cancer cells to the chemotherapeutic drug oxaliplatin (OXA) by targeting YBX1. Cancer Biomark. 18:1–9. 2017.PubMed/NCBI View Article : Google Scholar
25	Mlakar V, Berginc G, Volavsek M, Stor Z, Rems M and Glavac D: Presence of activating KRAS mutations correlates significantly with expression of tumour suppressor genes DCN and TPM1 in colorectal cancer. BMC Cancer. 9(282)2009.PubMed/NCBI View Article : Google Scholar