miR‑29b suppresses proliferation and induces apoptosis of hepatocellular carcinoma ascites H22 cells via regulating TGF‑β1 and p53 signaling pathway

Liu,Yan-Lu; Yang,Wen-Hao; Chen,Bao-Yi; Nie,Juan; Su,Zi-Ren; Zheng,Jing-Na; Gong,Shi-Ting; Chen,Jian-Nan; Jiang,Dongbo; Li,Yucui

doi:10.3892/ijmm.2021.4990

miR‑29b suppresses proliferation and induces apoptosis of hepatocellular carcinoma ascites H22 cells via regulating TGF‑β1 and p53 signaling pathway

Authors:
- Yan-Lu Liu
- Wen-Hao Yang
- Bao-Yi Chen
- Juan Nie
- Zi-Ren Su
- Jing-Na Zheng
- Shi-Ting Gong
- Jian-Nan Chen
- Dongbo Jiang
- Yucui Li
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Affiliations: School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, P.R. China, School of Pharmaceutical Sciences, Guangzhou Medical University, Guangzhou, Guangdong 510006, P.R. China, Department of Pharmacy, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524000, P.R. China
Published online on: June 25, 2021 https://doi.org/10.3892/ijmm.2021.4990
Article Number: 157
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

MicroRNA (miR)‑29b is a key tumor regulator. It can inhibit tumor cell proliferation, induce apoptosis, suppress tumor invasion and migration, thus delaying tumor progression. Our previous studies revealed an increased level of miR‑29b in hepatoma 22 (H22) cells in ascites tumor‑bearing mice. The present study investigated the effect of miR‑29b on proliferation and apoptosis of hepatocellular carcinoma ascites H22 cells and its association with the transforming growth factor‑β1 (TGF‑β1) signaling pathway and p53‑mediated apoptotic pathway. Briefly, H22 cells were transfected with miR‑29b‑3p (hereinafter referred to as miR‑29b) mimic or miR‑29b inhibitor. MTS cell proliferation assay and flow cytometry were used to analyze cell viability and apoptosis. The expression change of the TGF‑β1 signaling pathway and p53‑mediated apoptotic pathway were detected by reverse transcription‑quantitative PCR, western blotting and immunofluorescence. Furthermore, cells were treated with exogenous TGF‑β1 and TGF‑β1 small interfering RNA to evaluate the crosstalk between TGF‑β1 and p53 under miR‑29b regulation. The overexpression of miR‑29b decreased cell viability, increased cell apoptosis, activated the TGF‑β1 signaling pathway and p53‑mediated apoptotic pathway. Conversely, these effects were reversed by the miR‑29b inhibitor. Moreover, the effect of miR‑29b mimic was further increased after treating cells with exogenous TGF‑β1. The activation of the TGF‑β1 signaling pathway and p53‑mediated apoptotic pathway induced by miR‑29b overexpression were reversed by TGF‑β1 inhibition. In summary, these data indicated that miR‑29b has an important role in proliferation and apoptosis of H22 cells by regulating the TGF‑β1 signaling pathway, the p53‑dependent apoptotic pathway, and the crosstalk between TGF‑β1 and p53.

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1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	You LN, Tai QW, Xu L, Hao Y, Guo WJ, Zhang Q, Tong Q, Zhang H and Huang WK: Exosomal LINC00161 promotes angiogenesis and metastasis via regulating miR-590-3p/ROCK axis in hepatocellular carcinoma. Cancer Gene Ther. 28:719–736. 2021. View Article : Google Scholar : PubMed/NCBI
3	Smith EM and Jayson GC: The current and future management of malignant ascites. Clin Oncol (R Coll Radiol). 15:59–72. 2003. View Article : Google Scholar
4	Mendell JT and Olson EN: MicroRNAs in stress signaling and human disease. Cell. 148:1172–1187. 2012. View Article : Google Scholar : PubMed/NCBI
5	Croce CM: 37 causes and consequences of microRNA dysregulation in cancer. Eur J Cancer. 48(Suppl 5): S8–S9. 2012. View Article : Google Scholar
6	Yan B, Guo Q, Fu FJ, Wang Z, Yin Z, Wei YB and Yang JR: The role of miR-29b in cancer: Regulation, function, and signaling. Onco Targets Ther. 8:539–548. 2015.PubMed/NCBI
7	Wang LH, Huang J, Wu CR, Huang LY, Cui J, Xing ZZ and Zhao CY: Downregulation of miR-29b targets DNMT3b to suppress cellular apoptosis and enhance proliferation in pancreatic cancer. Mol Med Rep. 17:2113–2120. 2018.
8	Cui H, Wang L, Gong P, Zhao C, Zhang S, Zhang K, Zhou R, Zhao Z and Fan H: Deregulation between miR-29b/c and DNMT3A is associated with epigenetic silencing of the CDH1 gene, affecting cell migration and invasion in gastric cancer. PLoS One. 10:e01239262015. View Article : Google Scholar : PubMed/NCBI
9	Wang B, Li W, Liu H, Yang L, Liao Q, Cui S, Wang H and Zhao L: miR-29b suppresses tumor growth and metastasis in colorectal cancer via downregulating Tiam1 expression and inhibiting epithelial-mesenchymal transition. Cell Death Dis. 5:e13352014. View Article : Google Scholar : PubMed/NCBI
10	Chen B, Wang J, Wang J, Wang H, Gu X, Tang L and Feng X: A regulatory circuitry comprising TP53, miR-29 family, and SETDB1 in non-small cell lung cancer. Biosci Rep. 38:BSR201806782018. View Article : Google Scholar :
11	Xiong Y, Fang JH, Yun JP, Yang J, Zhang Y, Jia WH and Zhuang SM: Effects of microRNA-29 on apoptosis, tumorigenicity, and prognosis of hepatocellular carcinoma. Hepatology. 51:836–845. 2010.
12	Yang HM, Sun CY, Liang JL, Xu LQ, Zhang ZB, Luo DD, Chen HB, Huang YZ, Wang Q, Lee DY, et al: Supercritical-carbon dioxide fluid extract from chrysanthemum indicum enhances anti-tumor effect and reduces toxicity of bleomycin in tumor-bearing mice. Int J Mol Sci. 18:4652017. View Article : Google Scholar :
13	Nie J, Yang HM, Sun CY, Liu YL, Zhuo JY, Zhang ZB, Lai XP, Su ZR and Li YC: Scutellarin enhances antitumor effects and attenuates the toxicity of bleomycin in H22 ascites tumor-bearing mice. Front Pharmacol. 9:6152018. View Article : Google Scholar : PubMed/NCBI
14	He XX, Zhang YN, Yan JW, Yan JJ, Wu Q and Song YH: CP-31398 inhibits the growth of p53-mutated liver cancer cells in vitro and in vivo. Tumour Biol. 37:807–815. 2016. View Article : Google Scholar
15	Liu L, Yu ZY, Yu TT, Cui SH, Yang L, Chang H, Qu YH, Lv XF, Zhang XA and Ren CC: A Slug-dependent mechanism is responsible for tumor suppression of p53-stabilizing compound CP-31398 in p53-mutated endometrial carcinoma. J Cell Physiol. 235:8768–8778. 2020. View Article : Google Scholar : PubMed/NCBI
16	Guo H, Zhang Z, Su Z, Sun C, Zhang X, Zhao X, Lai X, Su Z, Li Y and Zhan JY: Enhanced anti-tumor activity and reduced toxicity by combination andrographolide and bleomycin in ascitic tumor-bearing mice. Eur J Pharmacol. 776:52–63. 2016. View Article : Google Scholar : PubMed/NCBI
17	Avasarala S, Van Scoyk M, Wang J, Sechler M, Vandervest K, Brzezinski C, Weekes C, Edwards MG, Arcaroli J, Davis RE, et al: Hsa-miR29b, a critical downstream target of non-canonical Wnt signaling, plays an anti-proliferative role in non-small cell lung cancer cells via targeting MDM2 expression. Biol Open. 2:675–685. 2013. View Article : Google Scholar : PubMed/NCBI
18	Bierie B and Moses HL: Tumour microenvironment: TGFbeta: The molecular Jekyll and Hyde of cancer. Nat Rev Cancer. 6:506–520. 2006. View Article : Google Scholar : PubMed/NCBI
19	Zhou H, Wang K, Hu Z and Wen J: TGF-β1 alters microRNA profile in human gastric cancer cells. Chin J Cancer Res. 25:102–111. 2013.PubMed/NCBI
20	Zhenye L, Chuzhong L, Youtu W, Xiaolei L, Lei C, Lichuan H, Hongyun W, Yonggang W, Fei W and Yazhuo Z: The expression of TGF-β1, Smad3, phospho-Smad3 and Smad7 is correlated with the development and invasion of nonfunctioning pituitary adenomas. J Transl Med. 12:712014. View Article : Google Scholar
21	Buenemann CL, Willy C, Buchmann A, Schmiechen A and Schwarz M: Transforming growth factor-beta1-induced Smad signaling, cell-cycle arrest and apoptosis in hepatoma cells. Carcinogenesis. 22:447–452. 2001. View Article : Google Scholar : PubMed/NCBI
22	Johansson J, Tabor V, Wikell A, Jalkanen S and Fuxe J: TGF-β1-induced epithelial-mesenchymal transition promotes monocyte/macrophage properties in breast cancer cells. Front Oncol. 5:32015. View Article : Google Scholar
23	Han G and Wang XJ: Roles of TGFβ signaling Smads in squamous cell carcinoma. Cell Biosci. 1:412011. View Article : Google Scholar
24	Zhang G, Feng W and Wu J: Down-regulation of SEPT9 inhibits glioma progression through suppressing TGF-β-induced epithelial-mesenchymal transition (EMT). Biomed Pharmacother. 125:1097682020. View Article : Google Scholar
25	Takahashi K, Menju T, Nishikawa S, Miyata R, Tanaka S, Yutaka Y, Yamada Y, Nakajima D, Hamaji M, Ohsumi A, et al: Tranilast inhibits TGF-β1-induced epithelial-mesenchymal transition and invasion/metastasis via the suppression of Smad4 in human lung cancer cell lines. Anticancer Res. 40:3287–3296. 2020. View Article : Google Scholar : PubMed/NCBI
26	Samarakoon R, Higgins SP, Higgins CE and Higgins PJ: The TGF-β1/p53/PAI-1 signaling axis in vascular senescence: Role of caveolin-1. Biomolecules. 9:3412019. View Article : Google Scholar
27	Saile B, Matthes N, El Armouche H, Neubauer K and Ramadori G: The bcl, NFkappaB and p53/p21WAF1 systems are involved in spontaneous apoptosis and in the anti-apoptotic effect of TGF-beta or TNF-alpha on activated hepatic stellate cells. Eur J Cell Biol. 80:554–561. 2001. View Article : Google Scholar : PubMed/NCBI
28	Sun Y, Xia P, Zhang H, Liu B and Shi Y: P53 is required for Doxorubicin-induced apoptosis via the TGF-beta signaling pathway in osteosarcoma-derived cells. Am J Cancer Res. 6:114–125. 2016.PubMed/NCBI
29	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. View Article : Google Scholar
30	Kim S, Lee JH, Kang I, Hyun S, Yu J and Shin C: An amphiphilic peptide induces apoptosis through the miR29b-p53 pathway in cancer cells. Mol Ther Nucleic Acids. 5:e3302016. View Article : Google Scholar : PubMed/NCBI
31	Rupaimoole R and Slack FJ: MicroRNA therapeutics: Towards a new era for the management of cancer and other diseases. Nat Rev Drug Discov. 16:203–222. 2017. View Article : Google Scholar : PubMed/NCBI
32	Tu HC, Jacobs SC, Borkowski A and Kyprianou N: Incidence of apoptosis and cell proliferation in prostate cancer: Relationship with TGF-beta1 and bcl-2 expression. Int J Cancer. 69:357–363. 1996. View Article : Google Scholar : PubMed/NCBI
33	Oltvai ZN, Milliman CL and Korsmeyer SJ: Bcl-2 heterodimerizes in vivo with a conserved homolog, bax, that accelerates programmed cell death. Cell. 74:609–619. 1993. View Article : Google Scholar : PubMed/NCBI
34	Fries KL, Miller WE and Raab-Traub N: Epstein-Barr virus latent membrane protein 1 blocks p53-mediated apoptosis through the induction of the A20 gene. J Virol. 70:8653–8659. 1996. View Article : Google Scholar : PubMed/NCBI
35	Miyashita T and Reed JC: Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell. 80:293–299. 1995. View Article : Google Scholar : PubMed/NCBI
36	Zhu S, Wang T, Luo F, Li H, Jia Q, He T, Wu H and Zou T: Astaxanthin inhibits proliferation and induces apoptosis of LX-2 cells by regulating the miR-29b/Bcl-2 pathway. Mol Med Rep. 19:3537–3547. 2019.PubMed/NCBI
37	Liang H, Wang Q, Wang D, Zheng H, Kalvakolanu DV, Lu H, Wen N, Chen X, Xu L, Ren J, et al: RGFP966, a histone deacetylase 3 inhibitor, promotes glioma stem cell differentiation by blocking TGF-β signaling via SMAD7. Biochem Pharmacol. 180:1141182020. View Article : Google Scholar
38	Li B, Yin GF, Wang YL, Tan YM, Huang CL and Fan XM: Impact of fecal microbiota transplantation on TGF-β1/Smads/ERK signaling pathway of endotoxic acute lung injury in rats. 3 Biotech. 10:522020. View Article : Google Scholar
39	Sivadas VP and Kannan S: The microRNA networks of TGFβ signaling in cancer. Tumour Biol. 35:2857–2869. 2014. View Article : Google Scholar
40	Aure MR, Jernstrom S, Krohn M; Due Oslo Breast Cancer Consortium E; Mills GB, Borresen-Dale AL, Sahlberg KK, Lingjaerde OC and Kristensen VN: 331: Integrative analysis reveals extensive association between microRNA expression and mRNA-protein translation. Eur J Cancer. 50(Suppl 5): S792014. View Article : Google Scholar