A novel colon cancer gene therapy using rAAV‑mediated expression of human shRNA-FHL2

Wu,Yao; Guo,Zheng; Zhang,Di; Zhang,Wenjing; Yan,Qingqing; Shi,Xinpeng; Zhang,Mengnan; Zhao,Yingying; Zhang,Yali; Jiang,Bo; Cheng,Tianming; Bai,Yang; Wang,Jide

doi:10.3892/ijo.2013.2090

A novel colon cancer gene therapy using rAAV‑mediated expression of human shRNA-FHL2

Authors:
- Yao Wu
- Zheng Guo
- Di Zhang
- Wenjing Zhang
- Qingqing Yan
- Xinpeng Shi
- Mengnan Zhang
- Yingying Zhao
- Yali Zhang
- Bo Jiang
- Tianming Cheng
- Yang Bai
- Jide Wang
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Affiliations: Guangdong Provincial Key Laboratory of Gastroenterology, Department of Digestive Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, P.R. China, Department of Oncology, The 1st Affiliated Hospital of Gannan Medical University, Ganzhou 341000, P.R. China, Department of Gastroenterology, The 1st Affiliated Hospital of Guangzhou Medical University, Guangzhou 510210, P.R. China
Published online on: September 5, 2013 https://doi.org/10.3892/ijo.2013.2090
Pages: 1618-1626

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Abstract

FHL2 (Four and a half LIM-only protein 2) has been identified as an oncogene in colon cancer and suppression of FHL2 induces cell differentiation and tumorigenesis in colon cancer cell lines. The aim of this study was to develop a novel and effective approach to knockdown FHL2, which can serve as a promising target of colon cancer therapy. Recombinant adeno-associated virus (rAAV) was generated bearing with FHL2-shRNA and transfected into LoVo cells. Cell cycle and growth were assessed. The interaction between FHL2 and G0/G1 cell cycle and growth was evaluated by flow cytometry, western blot analysis and WST-1 assay. We showed that suppression of FHL2 by rAAV-shRNA induced G0/G1 cell cycle arrest and inhibited cell growth. Apoptosis‑related proteins and their activity was investigated at the same time. rAAV-FHL2‑shRNA activated intrinsic and extrinsic apoptotic pathways and increased cell susceptibility to apoptotic stimuli by 5-FU. Moreover, a xenograft model was established to explore rAAV-FHL2-shRNA with 5-FU mediated tumorigenesis in vivo. A strong anti-tumorigenic effect of rAAV-FHL2-shRNA was shown in nude mice and this antitumor effect was enhanced when combined with 5-FU treatment. These findings implicate FHL2 as a cell cycle and growth modulator and thus inhibit apoptosis in colon cancer cells. rAAV-shRNA-FHL2 may serve as a novel and potent therapeutic or 5-FU co-therapeutic agent for colon cancer.

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