Effect of beclin 1 expression on the biological behavior and chemotherapy sensitivity of cervical cancer cells

Zhang,Yongxin; Lin,Shuang; Zhang,Yue; Chang,Suwen

doi:10.3892/ol.2016.4542

Effect of beclin 1 expression on the biological behavior and chemotherapy sensitivity of cervical cancer cells

Authors:
- Yongxin Zhang
- Shuang Lin
- Yue Zhang
- Suwen Chang
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Affiliations: Department of Gynecology, Yantai Shan Hospital, Yantai, Shandong 264000, P.R. China, Department of Gynecology, Yantai Yuhuangding Hospital Affiliated to Qingdao University, Yantai, Shandong 264000, P.R. China
Published online on: May 6, 2016 https://doi.org/10.3892/ol.2016.4542
Pages: 4089-4094

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Abstract

The present study aimed to evaluate the effect of the expression of the autophagic gene beclin 1 on the biological behavior and chemotherapy sensitivity towards Taxol® of cervical cancer HeLa cells. A beclin 1 expression vector was constructed and tranfected into HeLa cells. Reverse transcription‑polymerase chain reaction and western blotting were used to detect the expression of beclin 1. Cell proliferation was detected based on the growth curve of the cells. The effect of beclin 1 expression on cell apoptosis was analyzed using Hoechst 33258 staining, which enabled to observe the morphology of apoptotic cells. Apoptosis‑associated proteins were measured by western blot assay. The sensitivity of HeLa cells to Taxol® was analyzed by 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay. Beclin 1 expression at the messenger RNA and protein levels was elevated following transfection of the beclin 1 expression plasmid (P<0.05). Hoechst 33258 staining revealed that the apoptosis rate of the transfected HeLa cells was significantly higher than that of normal HeLa cells. The expression of caspase‑3 was increased in the transfected cells, and beclin 1 transfection increased B‑cell lymphoma 2 (Bcl‑2)‑associated X protein (Bax):Bcl‑2 ratio, resulting in Bax activation and Bcl‑2 suppression (P<0.05). Chemotherapy sensitivity analysis demonstrated that the half maximal inhibitory concentration values of Taxol® of the transfection, non‑transfection and mock‑vehicle groups were 30.4, 118.0 and 125.5 µg/ml, respectively. Beclin 1 inhibited proliferation and increased apoptosis of HeLa cells, and also increased the chemosensitivity of these cells to Taxol®. The present results confirmed that beclin 1 is a favorable prognostic biomarker for cervical cancer treatment, and may serve to identify particular patients for individual therapy.

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1	Jia Y, Li S, Yang R, Zhou H, Xiang Q, Hu T, Zhang Q, Chen Z, Ma D and Feng L: Knowledge about cervical cancer and barriers of screening program among women in Wufeng County, a high-incidence region of cervical cancer in China. PLoS One. 8:e670052013. View Article : Google Scholar : PubMed/NCBI
2	Sun Y, Liu JH, Sui YX, Jin L, Yang Y, Lin SM and Shi H: Beclin1 overexpression inhibitis proliferation, invasion and migration of CaSki cervical cancer cells. Asian Pac J Cancer Prev. 12:1269–1273. 2011.PubMed/NCBI
3	Zhang SK, Kang LN, Chang IJ, Zhao FH, Hu SY, Chen W, Shi JF, Zhang X, Pan QJ, Li SM and Qiao YL: The natural history of cervical cancer in Chinese women: Results from an 11-year follow-up study in China using a multistate model. Cancer Epidemiol Biomarkers Prev. 23:1298–1305. 2014. View Article : Google Scholar : PubMed/NCBI
4	Deng Q, Wang Z, Wang L, Zhang L, Xiang X, Wang Z and Chong T: Lower mRNA and protein expression levels of LC3 and Beclin1, markers of autophagy, were correlated with progression of renal clear cell carcinoma. Jpn J Clin Oncol. 43:1261–1268. 2013. View Article : Google Scholar : PubMed/NCBI
5	Wang ZH, Li L, Peng ZL and Duan ZL: Effect of autophagy gene Beclin 1 on the growth of cervical cancer HeLa cells in vitro and in vivo. Zhonghua Zhong Liu Za Zhi. 33:804–809. 2011.(In Chinese). PubMed/NCBI
6	Denton D, Nicolson S and Kumar S: Cell death by autophagy: Facts and apparent artefacts. Cell Death Differ. 19:87–95. 2012. View Article : Google Scholar : PubMed/NCBI
7	Liang XH, Kleeman LK, Jiang HH, Gordon G, Goldman JE, Berry G, Herman B and Levine B: Protection against fatal Sindbis virus encephalitis by beclin, a novel Bcl-2-interacting protein. J Virol. 72:8586–8596. 1998.PubMed/NCBI
8	Oberstein A, Jeffrey PD and Shi Y: Crystal structure of the Bcl-XL-Beclin 1 peptide complex: Beclin 1 is a novel BH3-only protein. J Biol Chem. 282:13123–13132. 2007. View Article : Google Scholar : PubMed/NCBI
9	Liu B, Bao JK, Yang JM and Cheng Y: Targeting autophagic pathways for cancer drug discovery. Chin J Cancer. 32:113–20. 2013. View Article : Google Scholar : PubMed/NCBI
10	Meijer AJ and Codogno P: Regulation and role of autophagy in mammalian cells. Int J Biochem Cell Biol. 36:2445–2462. 2004. View Article : Google Scholar : PubMed/NCBI
11	Sun Y, Liu JH, Jin L, Lin SM, Yang Y, Sui YX and Shi H: Over-expression of the Beclin1 gene upregulates chemosensitivity to anti-cancer drugs by enhancing therapy-induced apoptosis in cervix squamous carcinoma CaSki cells. Cancer Lett. 294:204–210. 2010. View Article : Google Scholar : PubMed/NCBI
12	Jin S and White E: Role of autophagy in cancer: Management of metabolic stress. Autophagy. 3:28–31. 2007. View Article : Google Scholar : PubMed/NCBI
13	Shin JY, Hong SH, Kang B, Minai-Tehrani A and Cho MH: Overexpression of beclin1 induced autophagy and apoptosis in lungs of K-rasLA1 mice. Lung Cancer. 81:362–370. 2013. View Article : Google Scholar : PubMed/NCBI
14	He C and Klionsky DJ: Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet. 43:67–93. 2009. View Article : Google Scholar : PubMed/NCBI
15	Levine B and Kroemer G: Autophagy in the pathogenesis of disease. Cell. 132:27–42. 2008. View Article : Google Scholar : PubMed/NCBI
16	Kroemer G, Mariño G and Levine B: Autophagy and the integrated stress response. Mol Cell. 40:280–293. 2010. View Article : Google Scholar : PubMed/NCBI
17	Cheng Y, Ren X, Hait WN and Yang JM: Therapeutic targeting of autophagy in disease: Biology and pharmacology. Pharmacol Rev. 65:1162–1197. 2013. View Article : Google Scholar : PubMed/NCBI
18	Kimmelman AC: The dynamic nature of autophagy in cancer. Genes Dev. 25:1999–2010. 2011. View Article : Google Scholar : PubMed/NCBI
19	Ravikumar B, Sarkar S, Davies JE, Futter M, Garcia-Arencibia M, Green-Thompson ZW, Jimenez-Sanchez M, Korolchuk VI, Lichtenberg M, Luo S, et al: Regulation of mammalian autophagy in physiology and pathophysiology. Physiol Rev. 90:1383–1435. 2010. View Article : Google Scholar : PubMed/NCBI
20	Ambjørn M, Ejlerskov P, Liu Y, Lees M, Jäättelä M and Issazadeh-Navikas S: IFNB1/interferon-β-induced autophagy in MCF-7 breast cancer cells counteracts its proapoptotic function. Autophagy. 9:287–302. 2013. View Article : Google Scholar : PubMed/NCBI
21	Filippi-Chiela EC, Villodre ES, Zamin LL and Lenz G: Autophagy interplay with apoptosis and cell cycle regulation in the growth inhibiting effect of resveratrol in glioma cells. PLoS One. 6:e208492011. View Article : Google Scholar : PubMed/NCBI
22	Wirawan E, Lippens S, Vanden Berghe T, Romagnoli A, Fimia GM, Piacentini M and Vandenabeele P: Beclin1: A role in membrane dynamics and beyond. Autophagy. 8:6–17. 2012. View Article : Google Scholar : PubMed/NCBI
23	Li X, Yan J, Wang L, Xiao F, Yang Y, Guo X and Wang H: Beclin1 inhibition promotes autophagy and decreases gemcitabine-induced apoptosis in Miapaca2 pancreatic cancer cells. Cancer Cell Int. 13:262013. View Article : Google Scholar : PubMed/NCBI
24	Zhao J, Wang ML, Li Z, Gao DM, Cai Y, Chang J and Wang SP: Interferon-alpha-2b induces autophagy in hepatocellular carcinoma cells through Beclin1 pathway. Cancer Biol Med. 11:64–68. 2014.PubMed/NCBI
25	Duan ZL, Peng ZL, Wang ZH and Yan NH: Correlation of autophagy gene Beclin1 to tumorigenesis and development of epithelial ovarian cancer. Ai Zheng. 26:258–263. 2007.(In Chinese). PubMed/NCBI
26	Liang XH, Jackson S, Seaman M, Brown K, Kempkes B, Hibshoosh H and Levine B: Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature. 402:672–676. 1999. View Article : Google Scholar : PubMed/NCBI
27	Wu XY, Chen J, Cao QH, Dong M, Lin Q, Fan XJ, Xia Q, Chen ZH, Liu Q and Wan XB: Beclin 1 activation enhances chemosensitivity and predicts a favorable outcome for primary duodenal adenocarcinoma. Tumour Biol. 34:713–722. 2013. View Article : Google Scholar : PubMed/NCBI
28	Wang TT, Cao QH, Chen MY, Xia Q, Fan XJ, Ma XK, Lin Q, Jia CC, Dong M, Ruan DY, et al: Beclin 1 deficiency correlated with lymph node metastasis, predicts a distinct outcome in intrahepatic and extrahepatic cholangiocarcinoma. PLoS One. 8:e803172013. View Article : Google Scholar : PubMed/NCBI
29	Sun Y, Jin L, Liu J, Lin S, Yang Y, Sui Y and Shi H: Effect of autophagy on paclitaxel-induced CaSki cell death. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 35:557–565. 2010.PubMed/NCBI
30	Niu TK, Cheng Y, Ren X and Yang JM: Interaction of Beclin 1 with survivin regulates sensitivity of human glioma cells to TRAIL-induced apoptosis. FEBS Lett. 584:3519–3524. 2010. View Article : Google Scholar : PubMed/NCBI
31	Lindsay J, Esposti MD and Gilmore AP: Bcl-2 proteins and mitochondria - specificity in membrane targeting for death. Biochim Biophys Acta. 1813:532–539. 2011. View Article : Google Scholar : PubMed/NCBI
32	Du P, Cao H, Wu HR, Zhu BS, Wang HW, Gu CW, Xing CG and Chen W: Blocking Bcl-2 leads to autophagy activation and cell death of the HEPG2 liver cancer cell line. Asian Pac J Cancer Prev. 14:5849–5854. 2013. View Article : Google Scholar : PubMed/NCBI
33	Yao Q, Chen J, Lv Y, Wang T, Zhang J, Fan J and Wang L: The significance of expression of autophagy-related gene Beclin, Bcl-2, and Bax in breast cancer tissues. Tumour Biol. 32:1163–1171. 2011. View Article : Google Scholar : PubMed/NCBI
34	Wang W, Fan H, Zhou Y, Duan P, Zhao G and Wu G: Knockdown of autophagy-related gene Beclin1 promotes cell growth and inhibits apoptosis in the A549 human lung cancer cell line. Mol Med Rep. 7:1501–1505. 2013.PubMed/NCBI