Synergistic inhibitory effects on hepatocellular carcinoma with recombinant human adenovirus Aspp2 and oxaliplatin via p53-independent pathway in vitro and in vivo

Liu,Xiaoni; Xu,Jianji; Wang,Shuang; Yu,Xiaoxiao; Kou,Boxin; Chai,Mengyin; Zang,Yunjin; Chen,Dexi

doi:10.3892/ijo.2017.4105

Synergistic inhibitory effects on hepatocellular carcinoma with recombinant human adenovirus Aspp2 and oxaliplatin via p53-independent pathway in vitro and in vivo

Authors:
- Xiaoni Liu
- Jianji Xu
- Shuang Wang
- Xiaoxiao Yu
- Boxin Kou
- Mengyin Chai
- Yunjin Zang
- Dexi Chen
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Affiliations: Beijing Institute of Hepatology and Beijing YouAn Hospital, Capital Medical University, Beijing 100069, P.R. China
Published online on: August 29, 2017 https://doi.org/10.3892/ijo.2017.4105
Pages: 1291-1299

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Abstract

The present study was designed to investigate the synergistic inhibitory effects on hepatocellular carcinoma with recombinant human adenovirus Aspp2 (Aspp2-ad) and oxaliplatin via p53-independent pathway in vitro and in vivo. After being treated with Aspp2-ad and/or oxaliplatin for 24-48 h, HepG2P53-/- and Hep3B cells showed a significant growth inhibition compared with vehicle control. Combination group showed a synergetic effect, the inhibitory rates were all above 80% at 48 h point in HepG2P53-/- and Hep3B cells. The apoptotic cell numbers of Aspp2-ad and/or oxaliplatin treatment groups were increased remarkably, especially for the combined therapy group in the liver cancer cells. The Hep3B xenograft experiment also showed similar inhibition of Aspp2-ad and/or oxaliplatin to the in vitro experiment. H&E results showed that combination group had the least mitotic indexes and the most necrosis. The immunohistochemistry results showed that PCNA, CD31 expression decreased greatly in treatment groups. These results suggested that Aspp2-ad might inhibit proliferation and vascular growth of hepatocarcinoma. Aspp2 induced apoptosis protein expression in Aspp2-ad and combination groups, the Aspp2, Bax and activation of caspase-3 expression increased greatly both in vitro and in vivo. But interestingly, the autophagy proteins showed different responses not only in HepG2P53-/- and Hep3B cells but also in vitro and in vivo. We found that Aspp2-ad downregulated the p-ERK, p-STAT3 expression, the synergistic effects were observed in combination group, while there was not response of mTOR to Aspp2-ad. In conclusion, Aspp2-ad, in P53-independent manner, regulated ERK and STAT3 signal moleculars to inhibit hepatocarcinoma in coordination with oxaliplatin by influencing the protein expression of proliferation, apoptosis, autophagy and vascular growth. Aspp2-ad has the potential to be developed in gene therapy for HCC, especially for P53 deletion or mutation in HCC.

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1	Lamarca A, Mendiola M and Barriuso J: Hepatocellular carcinoma: Exploring the impact of ethnicity on molecular biology. Crit Rev Oncol Hematol. 105:65–72. 2016. View Article : Google Scholar : PubMed/NCBI
2	Connell LC, Harding JJ and Abou-Alfa GK: Advanced hepatocellular cancer: The currentstate of future research. Curr Treat Options Oncol. 17:432016. View Article : Google Scholar
3	Grandhi MS, Kim AK, Ronnekleiv-Kelly SM, Kamel IR, Ghasebeh MA and Pawlik TM: Hepatocellular carcinoma: From diagnosis to treatment. Surg Oncol. 25:74–85. 2016. View Article : Google Scholar : PubMed/NCBI
4	Zhao C, Fan L, Qi F, Ou S, Yu L, Yi X, Ni B, Zheng Z, Lu J, Zhang C, et al: Raltitrexed plus oxaliplatin-based transarterial chemoembolization in patients with unresectable hepatocellular carcinoma. Anticancer Drugs. 27:689–694. 2016. View Article : Google Scholar : PubMed/NCBI
5	Vijayvergia N, Li T, Wong YN, Hall MJ, Cohen SJ and Dotan E: Chemotherapy use and adoption of new agents is affected by age and comorbidities in patients with metastatic colorectal cancer. Cancer. 122:3191–3198. 2016. View Article : Google Scholar : PubMed/NCBI
6	Ihara K, Yamaguchi S, Ueno N, Tani Y, Shida Y, Ogata H, Domeki Y, Okamoto K, Nakajima M, Sasaki K, et al: Expression of DNA double-strand break repair proteins predicts the response and prognosis of colorectal cancer patients undergoing oxaliplatin-based chemotherapy. Oncol Rep. 35:1349–1355. 2016. View Article : Google Scholar
7	Zhao C, Fan L, Qi F, Ou S, Yu L, Yi X, Ni B, Zheng Z, Lu J, Zhang C, et al: Raltitrexed plus oxaliplatin-based transarterial chemoembolization in patients with unresectable hepatocellular carcinoma. Anticancer Drugs. 27:689–694. 2016. View Article : Google Scholar : PubMed/NCBI
8	Gao S, Zhang PJ, Guo JH, Chen H, Xu HF, Liu P, Yang RJ and Zhu X: Chemoembolization alone vs combined chemoembolization and hepatic arterial infusion chemotherapy in inoperable hepatocellular carcinoma patients. World J Gastroenterol. 21:10443–10452. 2015. View Article : Google Scholar : PubMed/NCBI
9	Liu L, Zheng YH, Han L and Qin SK: Efficacy and safety of the oxaliplatin-based chemotherapy in the treatment of advanced primary hepatocellular carcinoma: A meta-analysis of prospective studies. Medicine (Baltimore). 95:e49932016. View Article : Google Scholar
10	Xie YS, Zhang YH, Liu SP, Liu SQ, Peng CW, Wu L, Luo HS and Li Y: Synergistic gastric cancer inhibition by chemogenetherapy with recombinant human adenovirus p53 and epirubicin: An in vitro and in vivo study. Oncol Rep. 24:1613–1620. 2010.PubMed/NCBI
11	Chen S, Chen J, Xi W, Xu W and Yin G: Clinical therapeutic effect and biological monitoring of p53 gene in advanced hepatocellular carcinoma. Am J Clin Oncol. 37:24–29. 2014. View Article : Google Scholar : PubMed/NCBI
12	Sangro B, Mazzolini G, Ruiz M, Ruiz J, Quiroga J, Herrero I, Qian C, Benito A, Larrache J, Olagüe C, et al: A phase I clinical trial of thymidine kinase-based gene therapy in advanced hepatocellular carcinoma. Cancer Gene Ther. 17:837–843. 2010. View Article : Google Scholar : PubMed/NCBI
13	Liu ZJ, Lu X, Zhang Y, Zhong S, Gu SZ, Zhang XB, Yang X and Xin HM: Downregulated mRNA expression of ASPP and the hypermethylation of the 5′-untranslated region in cancer cell lines retaining wild-type p53. FEBS Lett. 579:1587–1590. 2005. View Article : Google Scholar : PubMed/NCBI
14	Mori T, Okamoto H, Takahashi N, Ueda R and Okamoto T: Aberrant overexpression of 53BP2 mRNA in lung cancer cell lines. FEBS Lett. 465:124–128. 2000. View Article : Google Scholar : PubMed/NCBI
15	Liu WK, Jiang XY, Ren JK and Zhang ZX: Expression pattern of the ASPP family members in endometrial endometrioid adenocarcinoma. Onkologie. 33:500–503. 2010. View Article : Google Scholar : PubMed/NCBI
16	Mak VC, Lee L, Siu MK, Wong OG, Lu X, Ngan HY, Wong ES and Cheung AN: Downregulation of ASPP2 in choriocarcinoma contributes to increased migratory potential through Src signaling pathway activation. Carcinogenesis. 34:2170–2177. 2013. View Article : Google Scholar : PubMed/NCBI
17	Schittenhelm MM, Illing B, Ahmut F, Rasp KH, Blumenstock G, Döhner K, Lopez CD and Kampa-Schittenhelm KM: Attenuated expression of apoptosis stimulating protein of p53-2 (ASPP2) in human acute leukemia is associated with therapy failure. PLoS One. 8:e801932013. View Article : Google Scholar : PubMed/NCBI
18	Li Y, Li B, Li CJ and Li LJ: Key points of basic theories and clinical practice in rAd-p53 (Gendicine™) gene therapy for solid malignant tumors. Expert Opin Biol Ther. 15:437–454. 2015. View Article : Google Scholar
19	Chen GX, Zhang S, He XH, Liu SY, Ma C and Zou XP: Clinical utility of recombinant adenoviral human p53 gene therapy: Current perspectives. Onco Targets Ther. 7:1901–1909. 2014. View Article : Google Scholar : PubMed/NCBI
20	Shi Y, Han Y, Xie F, Wang A, Feng X, Li N, Guo H and Chen D: ASPP2 enhances oxaliplatin (L-OHP)-induced colorectal cancer cell apoptosis in a p53-independent manner by inhibiting cell autophagy. J Cell Mol Med. 19:535–543. 2015. View Article : Google Scholar :
21	Wang S, XU JJ, Liu XN and Chen DX: Quality control of human ASPP2 recombinant adenovirus, 2016. Zhongguo Yao Li Xue Tong Bao. 32:885–888. 2016.In Chinese.
22	Amer MH: Gene therapy for cancer: Present status and future perspective. Mol Cell Ther. 2:272014. View Article : Google Scholar : PubMed/NCBI
23	Sheridan C: Gene therapy finds its niche. Nat Biotechnol. 29:121–128. 2011. View Article : Google Scholar : PubMed/NCBI
24	Tazawa H, Kagawa S and Fujiwara T: Advances in adenovirus-mediated p53 cancer gene therapy. Expert Opin Biol Ther. 13:1569–1583. 2013. View Article : Google Scholar : PubMed/NCBI
25	Chiocca EA, Abbed KM, Tatter S, Louis DN, Hochberg FH, Barker F, Kracher J, Grossman SA, Fisher JD, Carson K, et al: A phase I open-label, dose-escalation, multi-institutional trial of injection with an E1B-Attenuated adenovirus, ONYX-015, into the peritumoral region of recurrent malignant gliomas, in the adjuvant setting. Mol Ther. 10:958–966. 2004. View Article : Google Scholar : PubMed/NCBI
26	Block SL, Nolan T, Sattler C, Barr E, Giacoletti KE, Marchant CD, Castellsagué X, Rusche SA, Lukac S, Bryan JT, et al Protocol 016 Study Group: Comparison of the immunogenicity and reactogenicity of a prophylactic quadrivalent human papillomavirus (types 6, 11, 16, and 18) L1 virus-like particle vaccine in male and female adolescents and young adult women. Pediatrics. 118:2135–2145. 2006. View Article : Google Scholar : PubMed/NCBI
27	Kantoff PW, Schuetz TJ, Blumenstein BA, Glode LM, Bilhartz DL, Wyand M, Manson K, Panicali DL, Laus R, Schlom J, et al: Overall survival analysis of a phase II randomized controlled trial of a Poxviral-based PSA-targeted immunotherapy in metastatic castration-resistant prostate cancer. J Clin Oncol. 28:1099–1105. 2010. View Article : Google Scholar : PubMed/NCBI
28	van Putten EH, Dirven CM, van den Bent MJ and Lamfers ML: Sitimagene ceradenovec: A gene-based drug for the treatment of operable high-grade glioma. Future Oncol. 6:1691–1710. 2010. View Article : Google Scholar : PubMed/NCBI
29	Liu J, Li W, Deng M, Liu D, Ma Q and Feng X: Immunohistochemical determination of p53 protein overexpression for predicting p53 gene mutations in hepatocellular carcinoma: A meta-analysis. PLoS One. 2016.11:e01596362016. View Article : Google Scholar : PubMed/NCBI
30	Liu J, Ma Q, Zhang M, Wang X, Zhang D, Li W, Wang F and Wu E: Alterations of TP53 are associated with a poor outcome for patients with hepatocellular carcinoma: Evidence from a systematic review and meta-analysis. Eur J Cancer. 48:2328–2338. 2012. View Article : Google Scholar : PubMed/NCBI
31	Qi LN, Bai T, Chen ZS, Wu FX, Chen YY, De Xiang B, Peng T, Han ZG and Li LQ: The p53 mutation spectrum in hepatocellular carcinoma from Guangxi, China: Role of chronic hepatitis B virus infection and aflatoxin B1 exposure. Liver Int. 35:999–1009. 2015. View Article : Google Scholar
32	Bravo R, Fey SJ, Bellatin J, Larsen PM, Arevalo J and Celis JE: Identification of a nuclear and of a cytoplasmic polypeptide whose relative proportions are sensitive to changes in the rate of cell proliferation. Exp Cell Res. 136:311–319. 1981. View Article : Google Scholar : PubMed/NCBI
33	Yin S, Li Z, Huang J, Miao Z, Zhang J, Lu C and Xu H and Xu H: Prognostic value and clinicopathological significance of proliferating cell nuclear antigen expression in gastric cancer: A systematic review and meta-analysis. Onco Targets Ther. 10:319–327. 2017. View Article : Google Scholar : PubMed/NCBI
34	Ma S, Yang J, Li J and Song J: The clinical utility of the proliferating cell nuclear antigen expression in patients with hepatocellular carcinoma. Tumour Biol. 37:7405–7412. 2016. View Article : Google Scholar
35	Pantanowitz L, Moses AV and Früh K: CD31 immunohistochemical staining in Kaposi Sarcoma. Arch Pathol Lab Med. 136:1329author reply 1330. 2012. View Article : Google Scholar : PubMed/NCBI
36	Delou JM, Biasoli D and Borges HL: The complex link between apoptosis and autophagy: A promising new role for RB. An Acad Bras Cienc. 88:2257–2275. 2016. View Article : Google Scholar : PubMed/NCBI
37	Ryter SW, Mizumura K and Choi AM: The impact of autophagy on cell death modalities. Int J Cell Biol. 2014:5026762014. View Article : Google Scholar : PubMed/NCBI
38	Su M, Mei Y and Sinha S: Role of the crosstalk between autophagy and apoptosis in cancer, 2013. J Oncol. 2013:1027352013. View Article : Google Scholar
39	Mariño G, Niso-Santano M, Baehrecke EH and Kroemer G: Self-consumption: The interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol. 15:81–94. 2014. View Article : Google Scholar : PubMed/NCBI
40	Sun Y, Liu WZ, Liu T, Feng X, Yang N and Zhou HF: Signaling pathway of MAPK/ERK in cell proliferation, differentiation, migration, senescence and apoptosis. J Recept Signal Transduct Res. 35:600–604. 2015. View Article : Google Scholar : PubMed/NCBI
41	Li L, Zhao GD, Shi Z, Qi LL, Zhou LY and Fu ZX: The Ras/Raf/MEK/ERK signaling pathway and its role in the occurrence and development of HCC. Oncol Lett. 12:3045–3050. 2016.PubMed/NCBI
42	Sarbassov DD, Ali SM and Sabatini DM: Growing roles for the mTOR pathway. Curr Opin Cell Biol. 17:596–603. 2005. View Article : Google Scholar : PubMed/NCBI
43	Cao Z, Liu LZ, Dixon DA, Zheng JZ, Chandran B and Jiang BH: Insulin-like growth factor-I induces cyclooxygenase-2 expression via PI3K.MAPK and PKC sign aling pathways in human ovarian cancer cells, 2007. Cel Signal. 19:1542–1553. 2007. View Article : Google Scholar
44	Matei D, Kelich S, Cao L, Menning N, Emerson RE, Rao J, Jeng MH and Sledge GW: PDGF BB induces VEGF secretion in ovarian cancer. Cancer Biol Ther. 6:1951–1959. 2007. View Article : Google Scholar : PubMed/NCBI
45	Park CM, Park MJ, Kwak HJ, Lee HC, Kim MS, Lee SH, Park IC, Rhee CH and Hong SI: Ionizing radiation enhances matrix metal-loproteinase-2 secretion and invasion of glioma cells through Src/epidermal growth factor receptor-mediated p38/Akt and phosphatidylinositol 3-kinase/Akt signaling pathways. Cancer Res. 66:8511–8519. 2006. View Article : Google Scholar : PubMed/NCBI
46	Zhou Q, Wong CH, Lau CP, Hui CW, Lui VW, Chan SL and Yeo W: Enhanced antitumor activity with combining effect of mTOR inhibition and microtubule stabilization in hepatocellular carcinoma. Int J Hepatol. 2013:1038302013. View Article : Google Scholar : PubMed/NCBI
47	Mali SB: Review of STAT3 (Signal Transducers and Activators of Transcription) in head and neck cancer. Oral Oncol. 51:565–569. 2015. View Article : Google Scholar : PubMed/NCBI
48	Li CJ, Li YC, Zhang DR and Pan JH: Signal transducers and activators of transcription 3 function in lung cancer. J Cancer Res Ther. 9(Suppl 2): S67–S73. 2013. View Article : Google Scholar : PubMed/NCBI
49	Subramaniam A, Shanmugam MK, Perumal E, Li F, Nachiyappan A, Dai X, Swamy SN, Ahn KS, Kumar AP, Tan BK, et al: Potential role of signal transducer and activator of transcription (STAT)3 signaling pathway in inflammation, survival, proliferation and invasion of hepatocellular carcinoma. Biochim Biophys Acta. 1835:46–60. 2013.