Hiwi overexpression does not affect proliferation, migration or apoptosis of liver cancer cells in vitro or in vivo

Li,Hao; Shi,Chen‑Xi; Liu,Hui; Zhang,Hai‑Han; Sang,Huai‑Ming; Soyfoo,Muhammad‑Djaleel; Cao,Jiu‑Liang; Xu,Shun‑Fu; Jiang,Jian‑Xia

doi:10.3892/ol.2018.8585

Hiwi overexpression does not affect proliferation, migration or apoptosis of liver cancer cells in vitro or in vivo

Authors:
- Hao Li
- Chen‑Xi Shi
- Hui Liu
- Hai‑Han Zhang
- Huai‑Ming Sang
- Muhammad‑Djaleel Soyfoo
- Jiu‑Liang Cao
- Shun‑Fu Xu
- Jian‑Xia Jiang
View Affiliations

Affiliations: Department of Pathophysiology, Nanjing Medical University, Nanjing, Jiangsu 211166, P.R. China, Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China
Published online on: April 26, 2018 https://doi.org/10.3892/ol.2018.8585
Pages: 9711-9718
Copyright: © Li 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

Piwi like RNA‑mediated gene silencing 1 (Hiwi) is a human homolog of the Piwi gene family that has been reported to be upregulated in hepatocellular carcinoma (HCC). The present study aimed to investigate the role of Hiwi in the initiation and development of HCC in vitro and in vivo. Adenovirus‑mediated Hiwi overexpression was established in primary murine hepatocytes and SMMC7721 HCC cells. Cell viability and proliferation were assessed using MTT and EdU assays, respectively. Cell migration was measured using a scratch migration assay. The cell cycle was assessed using flow cytometry, and the expression of genes associated with the epithelial mesenchymal transition (EMT) was assessed using reverse transcription‑quantitative polymerase chain reaction. SMMC7721 cells that stably express Hiwi were also generated and injected subcutaneously into the nude mice, and tumor growth was examined. Recombinant adenovirus encoding green fluorescent protein or Hiwi was delivered by injection into the tail vein, and its effect on murine hepatocyte gene expression was studied. The present study revealed that the overexpression of Hiwi did not affect the proliferation or migration of liver cancer cells and failed to suppress perifosine‑ or doxorubicin‑induced apoptosis in vitro. The tumors of mice that were injected with Hiwi‑expressing SMMC7721 cells were not significantly larger compared with mice that were injected with control SMMC7721 cells. Hiwi overexpression did not noticeably alter the expression of genes involved in EMT, either in vitro or in vivo. The results of the present study indicate that although expression of Hiwi is associated with HCC development and progression in the clinic, it does not act as an oncogene in liver cancer cells.

View Figures

View References

1	Suzuki R, Honda S and Kirino Y: PIWI expression and function in cancer. Front Genet. 3:2042012. View Article : Google Scholar : PubMed/NCBI
2	Qiao D, Zeeman AM, Deng W, Looijenga LH and Lin H: Molecular characterization of hiwi, a human member of the piwi gene family whose overexpression is correlated to seminomas. Oncogene. 21:3988–3999. 2002. View Article : Google Scholar : PubMed/NCBI
3	He W, Wang Z, Wang Q, Fan Q, Shou C, Wang J, Giercksky KE, Nesland JM and Suo Z: Expression of HIWI in human esophageal squamous cell carcinoma is significantly associated with poorer prognosis. BMC Cancer. 9:4262009. View Article : Google Scholar : PubMed/NCBI
4	Grochola LF, Greither T, Taubert H, Möller P, Knippschild U, Udelnow A, Henne-Bruns D and Würl P: The stem cell-associated Hiwi gene in human adenocarcinoma of the pancreas: Expression and risk of tumour-related death. Br J Cancer. 99:1083–1088. 2008. View Article : Google Scholar : PubMed/NCBI
5	Liu X, Sun Y, Guo J, Ma H, Li J, Dong B, Jin G, Zhang J, Wu J, Meng L and Shou C: Expression of hiwi gene in human gastric cancer was associated with proliferation of cancer cells. Int J Cancer. 118:1922–1929. 2006. View Article : Google Scholar : PubMed/NCBI
6	Raeisossadati R, Abbaszadegan MR, Moghbeli M, Tavassoli A, Kihara AH and Forghanifard MM: Aberrant expression of DPPA2 and HIWI genes in colorectal cancer and their impacts on poor prognosis. Tumour Biol. 35:5299–5305. 2014. View Article : Google Scholar : PubMed/NCBI
7	Taubert H, Greither T, Kaushal D, Würl P, Bache M, Bartel F, Kehlen A, Lautenschläger C, Harris L, Kraemer K, et al: Expression of the stem cell self-renewal gene Hiwi and risk of tumour-related death in patients with soft-tissue sarcoma. Oncogene. 26:1098–1100. 2007. View Article : Google Scholar : PubMed/NCBI
8	Chen Z, Che Q, He X, Wang F, Wang H, Zhu M, Sun J and Wan X: Stem cell protein Piwil1 endowed endometrial cancer cells with stem-like properties via inducing epithelial-mesenchymal transition. BMC Cancer. 15:8112015. View Article : Google Scholar : PubMed/NCBI
9	Wang DW, Wang ZH, Wang LL, Song Y and Zhang GZ: Overexpression of hiwi promotes growth of human breast cancer cells. Asian Pac J Cancer Prev. 15:7553–7558. 2014. View Article : Google Scholar : PubMed/NCBI
10	Liu W, Gao Q, Chen K, Xue X, Li M, Chen Q, Zhu G and Gao Y: Hiwi facilitates chemoresistance as a cancer stem cell marker in cervical cancer. Oncol Rep. 32:1853–1860. 2014. View Article : Google Scholar : PubMed/NCBI
11	Siddiqi S, Terry M and Matushansky I: Hiwi mediated tumorigenesis is associated with DNA hypermethylation. PLoS One. 7:e337112012. View Article : Google Scholar : PubMed/NCBI
12	Wang X, Tong X, Gao H, Yan X, Xu X, Sun S, Wang Q and Wang J: Silencing HIWI suppresses the growth, invasion and migration of glioma cells. Int J Oncol. 45:2385–2392. 2014. View Article : Google Scholar : PubMed/NCBI
13	Liang D, Fang Z, Dong M, Liang C, Xing C, Zhao J and Yang Y: Effect of RNA interference-related HiWi gene expression on the proliferation and apoptosis of lung cancer stem cells. Oncol Lett. 4:146–150. 2012. View Article : Google Scholar : PubMed/NCBI
14	Sharma AK, Nelson MC, Brandt JE, Wessman M, Mahmud N, Weller KP and Hoffman R: Human CD34(+) stem cells express the hiwi gene, a human homologue of the Drosophila gene piwi. Blood. 97:426–434. 2001. View Article : Google Scholar : PubMed/NCBI
15	Wang Y, Jiang Y, Ma N, Sang B, Hu X, Cong X and Liu Z: Overexpression of Hiwi inhibits the growth and migration of chronic myeloid leukemia cells. Cell Biochem Biophys. 73:117–124. 2015. View Article : Google Scholar : PubMed/NCBI
16	Kao JH and Chen DS: Changing disease burden of hepatocellular carcinoma in the Far East and Southeast Asia. Liver Int. 25:696–703. 2005. View Article : Google Scholar : PubMed/NCBI
17	Jiang J, Zhang H, Tang Q, Hao B and Shi R: Expression of HIWI in human hepatocellular carcinoma. Cell Biochem Biophys. 61:53–58. 2011. View Article : Google Scholar : PubMed/NCBI
18	Zhao YM, Zhou JM, Wang LR, He HW, Wang XL, Tao ZH, Sun HC, Wu WZ, Fan J, Tang ZY and Wang L: HIWI is associated with prognosis in patients with hepatocellular carcinoma after curative resection. Cancer. 118:2708–2717. 2012. View Article : Google Scholar : PubMed/NCBI
19	Xie Y, Yang Y, Ji D, Zhang D, Yao X and Zhang X: Hiwi downregulation, mediated by shRNA, reduces the proliferation and migration of human hepatocellular carcinoma cells. Mol Med Rep. 11:1455–1461. 2015. View Article : Google Scholar : PubMed/NCBI
20	Webber C, Gospodarowicz M, Sobin LH, Wittekind C, Greene FL, Mason MD, Compton C, Brierley J and Groome PA: Improving the TNM classification: Findings from a 10-year continuous literature review. Int J Cancer. 135:371–378. 2014. View Article : Google Scholar : PubMed/NCBI
21	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 : PubMed/NCBI
22	Fei HR, Chen G, Wang JM and Wang FZ: Perifosine induces cell cycle arrest and apoptosis in human hepatocellular carcinoma cell lines by blockade of Akt phosphorylation. Cytotechnology. 62:449–460. 2010. View Article : Google Scholar : PubMed/NCBI
23	Miyashita T and Reed JC: Bcl-2 oncoprotein blocks chemotherapy-induced apoptosis in a human leukemia cell line. Blood. 81:151–157. 1993.PubMed/NCBI
24	Mizutani H, Tada-Oikawa S, Hiraku Y, Kojima M and Kawanishi S: Mechanism of apoptosis induced by doxorubicin through the generation of hydrogen peroxide. Life Sci. 76:1439–1453. 2005. View Article : Google Scholar : PubMed/NCBI
25	Al-Janabi O, Wach S, Nolte E, Weigelt K, Rau TT, Stöhr C, Legal W, Schick S, Greither T, Hartmann A, et al: Piwi-like 1 and 4 gene transcript levels are associated with clinicopathological parameters in renal cell carcinomas. Biochim Biophys Acta. 1842:686–690. 2014. View Article : Google Scholar : PubMed/NCBI
26	Iliev R, Stanik M, Fedorko M, Poprach A, Vychytilova-Faltejskova P, Slaba K, Svoboda M, Fabian P, Pacik D, Dolezel J and Slaby O: Decreased expression levels of PIWIL1, PIWIL2, and PIWIL4 are associated with worse survival in renal cell carcinoma patients. Onco Targets Ther. 9:217–222. 2016.PubMed/NCBI