Antitumor Research of Recombinant Adenovirus Carrying mK5 gene Combined with Docetaxel on LNCaP Cell in vitro

Hu Yun¹, Fang Xianlong², Gu Jinfa^2,3, Yang Yuanyuan², Li Hui², Liu Xinyuan^1,2,3, Zhang Kangjian^2,3*

(¹Xinyuan Institute of Medicine and Biotechnology, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China; ²Shanghai Yuan Song Biological Technology, Shanghai 201401, China; ³Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China)

Abstract:

A recombinant adenovirus carrying the mK5 gene (OncoAd-mK5) was combined with docetaxel (DTX) for the evaluation of antitumor effect in prostatic cancer cell line LNCaP in vitro. CCK8 assay was used to respectively detect inhibitory effects of OncoAd-mK5, docetaxel ,the combination of OncoAd-mK5 and docetaxel on the growth of prostatic cancer cell lines. Morphological analysis was checked by microscope in prostatic cell line LNCaP treated by single OncoAd-mK5 or docetaxel only, or by combination. Hoechst 33258 staining and flow cytometry assay were performed to determine the apoptosis effects of single and combinational therapy on prostatic cell line LNCaP, respectively. Besides, the protein levels of E1A, mK5, and apoptosis related protein Caspase-8, XIAP and PARP were determined by Western blot. In addition, real-time quantitative PCR was used to analyze changes in vascular endothelial growth factor (VEGF) mRNA levels in the treated cells. The results illustrated that 4 MOI oncolytic adenovirus OncoAd-mK5 and 5 nmol/L docetaxel could synergistically inhibit the growth of prostate cancer cell line LNCaP, and the apoptosis effect of the combination treated group was more obvious than that of the single treatment. This new drug combination strategy provides a reference for the treatment of prostate cancer in clinical situations.

CSTR: 32200.14.cjcb.2019.05.0012