Adenovirus encoding XAF-1 and TNF‑α in the same open reading frame efficiently inhibits hepatocellular cancer cells

Li,Kai; Li,Xinhong; Wu,Zhongjun; Zheng,Liansheng; Cui,Yuqin; Wang,Jun; Huang,Yin; Yan,Zhihong

doi:10.3892/mmr.2016.5193

Adenovirus encoding XAF-1 and TNF‑α in the same open reading frame efficiently inhibits hepatocellular cancer cells

Authors:
- Kai Li
- Xinhong Li
- Zhongjun Wu
- Liansheng Zheng
- Yuqin Cui
- Jun Wang
- Yin Huang
- Zhihong Yan
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Affiliations: Department of Hepatobiliary Surgery, Hepatobiliary Treatment Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China, Department of Medicine, First Affiliated Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia 010050, P.R. China, Department of Surgical Oncology, Baotou Cancer Hospital, Baotou, Inner Mongolia 014030, P.R. China, Department of Oncology, Baotou Cancer Hospital, Baotou, Inner Mongolia 014030, P.R. China, Department of Pharmacy, Baotou Cancer Hospital, Baotou, Inner Mongolia 014030, P.R. China
Published online on: April 26, 2016 https://doi.org/10.3892/mmr.2016.5193
Pages: 5169-5176

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Abstract

X‑linked inhibitor of apoptosis (XIAP)‑associated factor 1 (XAF‑1), a tumor suppressor, is downregulated in most human malignant tumors. However, the tumor suppressive role of XAF‑1 in hepatocellular carcinoma (HCC) and its therapeutic value require further elucidation. The present study examined the expression of XAF‑1 at the mRNA and protein level in the HCC and paired peritumor tissue specimens, as well as in HCC cell lines and a normal liver cell line. A recombinant adenovirus which co‑expressed XAF‑1 and TNF‑α was then constructed, and its effects on the proliferation and colony formation ability of the MHCC97H HCC cell line were assessed using apoptosis induction, flow cytometry, trypan blue staining assay and a clonogenic assay. The results demonstrated that the expression of XAF‑1 was significantly reduced in HCC tissues compared with that in their matched peritumor specimens, and a significant correlation with the tumor size, stage and tumor ‑ nodes ‑ metastasis stage was identified. The reduced levels of XAF‑1 were further confirmed the HCC cell lines MHCC97L, HepG2 and MHCC97H compared with those in the L02 normal liver cell line. The recombinant adenovirus Ad‑XAF‑1&TNF‑α, which co‑expressed XAF‑1 and TNF‑α, was shown to efficiently express the two proteins at the mRNA and protein level. Furthermore, infection with Ad‑XAF‑1&TNF‑α synergistically induced apoptosis, reduced the proliferation and colony formation ability of MHCC97L cells to a significantly greater extent than overexpression of XAF‑1 or TNF‑α individually. To the best of our knowledge, the present study was the first to construct an adenovirus which co‑expressed XAF‑1 and TNF‑α in the same open reading frame and expressed them proportionally. As Ad‑XAF‑1&TNF‑α inhibited HCC cells with enhanced efficiency, it may be applicable for the treatment of HCC.

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