Effect of Targeted Interference with TAGLN Expression on Biological Behavior of HBV-Positive Hepatocellular Carcinoma Cells and Its Mechanisms

doi:10.13523/j.cb.20191102

China Biotechnology

2019, Vol. 39

Issue (11): 13-21 DOI: 10.13523/j.cb.20191102

Effect of Targeted Interference with TAGLN Expression on Biological Behavior of HBV-Positive Hepatocellular Carcinoma Cells and Its Mechanisms

XU Yan¹,LIU Zheng-yun^1,²,ZHANG Wan-ling¹,WANG Sheng-yu^1,²,WANG Huan^1,^2,^**(

)

1 Research Center for Medical and Biology,Zunyi Medical University, Zunyi 563099, China
2 Key Laboratory of Infectious Disease & Biosafety, Provincial Department of Education, Zunyi 563099, China;

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Abstract

Objective: To invest igate the effect of TAGLN on the biological behavior of HBV-positive hepatocellular carcinoma cell line HepG2.2.15 and its underlying mechanism.Methods: Immunohistochemistry and Western blot were used to detect the expression levels of TAGLN in HBV-positive and HBV-negative hepatocellular carcinoma tissues and cells; HepG2.2.15 cells were infected with TAGLN interfering lentiviruses, and stable expression cell lines were screened by purinomycin. Then transfection efficiency was confirmed by Western blot; CCK-8 and clone formation assay were used to detect the proliferation of HepG2.2.15 cells. While cell migration and invasion capabilities were determined by Transwell assay. The expression levels of PI3K, p-PI3K, AKT and p-AKT were examined by Western blot.Results: The expression of TAGLN in HBV-positive hepatocellular carcinoma tissues and cells was higher than that in HBV-negative hepatocellular carcinoma tissues and cells(P<0.01); Knockdown of TAGLN could effectively inhibit cell proliferation, migration and invasion (P<0.01), as well as down-regulate the expression of PI3K, AKT(P<0.01), p-PI3K and p-AKT (P<0.05).Conclusion: HBV infection can increase the expression of TAGLN in hepatocellular carcinoma. After interfering with TAGLN expression, the proliferation, cloning, migration and invasion of HepG2.2.15 cells decreased, which may be related to the decrease of PI3K and AKT expression.

Key words： Transgelin HCC HBV AKT PI3K

Received: 14 April 2019 Published: 17 December 2019

ZTFLH:

R373

Corresponding Authors: Huan WANG E-mail: wanghuan928@163.com

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	Yan XU
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	Sheng-yu WANG
	Huan WANG

Cite this article:

XU Yan,LIU Zheng-yun,ZHANG Wan-ling,WANG Sheng-yu,WANG Huan. Effect of Targeted Interference with TAGLN Expression on Biological Behavior of HBV-Positive Hepatocellular Carcinoma Cells and Its Mechanisms. China Biotechnology, 2019, 39(11): 13-21.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20191102 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I11/13

Fig.1 The expression of TAGLN in HBV-positive hepatocellular carcinoma was higher than that in HBV-negative hepatocellular carcinoma (a)The results of paraffin section of human hepatocellular carcinoma tissues by immunohistochemical staining (b) Statistical analysis of TAGLN immunohistochemistry results (c) The results of Western blot analysis (d) Statistic analysis of Western blot ** P< 0.01

Fig.2 Stable expression of TAGLN in shTAGLN1^#-2.15 and shTAGLN2^#-2.15 cell lines (a) The results of Western blot analysis (b) Statistic analysis of Western blot ** P< 0.01

Fig.3 Interference with TAGLN expression inhibits the proliferation of HepG2.2.15 cells (a) shTAGLN1^#-2.15 cell viability (b)shTAGLN2^#-2.15 cell viability (c) The results of colony formation assay (d) Statistic analysis of colony formation assay ** P<0.01

Fig.4 Interference with TAGLN expression inhibits migration and invasion of HepG2.2.15 cells (a) The results of transwell migration assay (200×) (b) Statistic analysis of transwell migration assay (c) The results of transwell invasion assay (200×) (d) Statistic analysis of transwell invasion assay ** P<0.01

Fig.5 Expression of PI3K and p-PI3K, AKT and p-AKT proteins in HepG2.2.15 cells (a) The results of Western blot analysis (b)-(e) The relative protein expression of AKT, p-AKT, PI3K and p-PI3K * P<0.05 and ** P< 0.01


[1]	Chen W, Zheng R, Baade P D , et al. Cancer statistics in China, 2015. CA Cancer J Clin, 2016,66(2):115-132. doi: 10.3322/caac.21338 pmid: 26808342

[2]	Forner A, Bruix J . Biomarkers for early diagnosis of hepatocellular carcinoma. The Lancet Oncology, 2012,13(8):750-751. doi: 10.1016/S1470-2045(12)70271-1 pmid: 22738800

[3]	吕桂帅, 陈磊, 王红阳 . 我国肝癌研究的现状与前景. 生命科学, 2015,27(3):237-248.

[3]	Lv G S, Chen L, Wang H Y . Current situation and prospect of hepatocellular carcinoma research in China .Life Sciences. 2015,27(3):237-248.

[4]	Lok A S ,McMahon B J,Jr Brown R S, et al. Antiviral therapy for chronic hepatitis B viral infection in adults: a systematic review and meta-analysis. Hepatology, 2016,63(1):284-306. doi: 10.1002/hep.28280 pmid: 26566246

[5]	Polaris Observatory Collaborators . Global prevalence, treatment, and prevention of hepatitis B virus infection in 2016: a modelling study. Lancet Gastroenterology & Hepatology, 2018,3(6):383-403. doi: 10.1016/S2468-1253(19)30362-0 pmid: 31836320

[6]	Lees-Miller J P, Heeley D H, Smillie L B . An abundant and novel protein of 22kDa (SM22) is widely distributed in smooth muscles.Purification from bovine aorta. Biochemical Journal, 1987,244(3):705-709. doi: 10.1042/bj2440705 pmid: 3446186

[7]	Lee E K, Han G Y, Park H W , et al. Transgelin promotes migration and invasion of cancer stem cells. Journal of Proteome Research, 2010,9(10):5108-5117. doi: 10.1021/pr100378z pmid: 20707403

[8]	Zhou L, Zhang R, Zhang L , et al. Upregulation of transgelin is an independent factor predictive of poor prognosis in patients with advanced pancreatic cancer. Cancer Science, 2013,104(4):423-430. doi: 10.1111/cas.12107 pmid: 23331552

[9]	Liu Z, Li G, Gou Y , et al. JS-K, a nitric oxide prodrug, induces DNA damage and apoptosis in HBV-positive hepatocellular carcinoma HepG2.2.15 cell. Biomed Pharmacother, 2017,92(8):989-997. doi: 10.1016/j.biopha.2017.05.141 pmid: 28605880

[10]	Bray F, Ferlay J , SoerjomataramI R L, et al. Global cancer statistics 2018: Globocan estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2018,68(6):394-424. doi: 10.3322/caac.21492 pmid: 30207593

[11]	Wang F S, Fan J G, Zhang Z , et al. The global burden of liver disease: the major impact of China. Hepatology, 2014,60(6):2099-2108. doi: 10.1002/hep.27406

[12]	Tat Trung N, Duong D C, Tong H V , et al. Optimisation of quantitative miRNA panels to consolidate the diagnostic surveillance of HBV-related hepatocellular carcinoma. PLoS One, 2018,13(4):1-17. doi: 10.1371/journal.pone.0196081 pmid: 29672637

[13]	Wang F S, Fan J G, Zhang Z , et al. The global burden of liver disease: the major impact of China. Hepatology, 2014,60(6):2099-2108. doi: 10.1002/hep.27406

[14]	Zhang X P, Jiang Y B, Zhong C Q , et al. PRMT1 promoted HCC growth and metastasis in vitro and in vivo via activating the STAT3 signalling pathway. Cellular Physiology and Biochemistry, 2018,47(4):1643-1654. doi: 10.1159/000490983 pmid: 29945155

[15]	Yu X, Tang W, Yang Y , et al. Long noncoding RNA NKILA enhances the anti-cancer effects of baicalein in hepatocellular carcinoma via the regulation of NF-kappa B signaling. Chem Biol Interact, 2018,285(4):48-58. doi: 10.1016/j.cbi.2018.02.027 pmid: 29481769

[16]	Wu T, Dong X, Yu D , et al. Natural product pectolinarigenin inhibits proliferation, induces apoptosis, and causes G2/M phase arrest of HCC via PI3K/AKT/mTOR/ERK signaling pathway. Onco Targets Ther, 2018,3(11):8633-8642. doi: 10.2147/OTT.S186186 pmid: 30584322

[17]	Buontempo F, Ersahin T, Missiroli S , et al. Inhibition of Akt signaling in hepatoma cells induces apoptotic cell death independent of Akt activation status. Invest New Drugs, 2011,29:1303-1313. doi: 10.1007/s10637-010-9486-3 pmid: 20628892

[18]	Ying D J, Ruan Y, Zhou X H , et al. MEG2 inhibits the growth and metastasis of hepatocellular carcinoma by inhibiting AKT pathway. Gene, 2019,687:1-8. doi: 10.1016/j.gene.2018.11.003 pmid: 30399427

[19]	Gdowski A, Panchoo M, Treuren T V , et al. Emerging therapeutics for targeting Akt in cancer. Front Biosci, 2016,21(1):757-768. doi: 10.1002/jcp.29333 pmid: 31663122

[20]	Kunter I, Erdal E, Nart D , et al. Active form of AKT controls cell proliferation and response to apoptosis in hepatocellular carcinoma. Oncology Reports, 2014,31(2):573-580. doi: 10.3892/or.2013.2932

[21]	Zhang C Z, Wang X D, Wang H W , et al. Sorafenib inhibits liver cancer growth by decreasing mTOR, AKT, and PI3K expression. Journal of Buon, 2015,20(1):218-222. pmid: 25778319

[22]	Mayer I A, Arteaga C L . The PI3K/AKT pathway as a target for cancer treatment. Annu Rev Med, 2016,67(1):11-28. doi: 10.1007/s11912-019-0846-7 pmid: 31828441

[23]	Courtney K D, Corcoran R B, Engelman J A . The PI3K pathway as drug target in human cancer. J Clin Oncol, 2010,28(6):1075-1083. doi: 10.1200/JCO.2009.25.3641 pmid: 20085938

[24]	Rawat S, Bouchard M J . The hepatitis B virus (HBV) HBx protein activates AKT to simultaneously regulate HBV replication and hepatocyte survival. Journal of Virology, 2015,89(2):999-1012. doi: 10.1128/JVI.02440-14 pmid: 25355887

[25]	Daniel C, Lüdke A, Wagner A , et al. Transgelin is a marker of repopulating mesangial cells after injury and promotes their proliferation and migration. Lab Invest, 2012,92(6):812-826. doi: 10.1038/labinvest.2012.63

[26]	Wu X, Dong L, Zhang R , et al. Transgelin overexpression in lung adenocarcinoma is associated with tumor progression. International Journal of Molecular Medicine, 2014,34(2):585-591. doi: 10.3892/ijmm.2014.1805

[27]	Kim T R, Cho E W, Paik S G , et al. Hypoxia-induced SM22 a in A549 cells activates the IGF1R/PI3K/Akt pathway,conferring cellular resistance against chemo- and radiation therapy. FEBS Lett, 2012,586(4):303-309. doi: 10.1016/j.febslet.2011.12.036

[28]	Zhou H, Zhang Y, Chen Q , et al. AKT and JNK signaling pathways increase the metastatic potential of colorectal cancer cells by altering transgelin expression. Digestive Diseases and Sciences, 2016,61(4):1091-1097. doi: 10.1007/s10620-015-3985-1 pmid: 26694173

[29]	白霞, 张连峰, 周琳 , 等. 靶向干扰骨架蛋白Transgelin抑制胰腺癌裸鼠移植瘤的生长. 中华普通外科杂志, 2013,28(7):538-541.

[29]	Bai X, Zhang L F, Zhou L , et al. Targeted interference with Transgelin inhibits the growth of transplanted pancreatic cancer in nude mice. Chinese Journal of General Surgery, 2013,28(7):538-541.

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