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Podophyllotoxin suppresses growth of MFC gastric cancer cells by modulating DNA damage response-related gene
Xuejing Li1, Yidan Pang1, Ya Liu1
1Department of Basic Medical Sciences, Medical College, Qinghai University, Xining, 810001 Qinghai Province P. R. China.
Abstract:
We first confirmed podophyllotoxin as the principal bioactive constituent in Qinghai-derived Sinopodophyllum hexandrum. Then, using the purified compound, we evaluated its inhibitory effects and mechanism in MFC gastric cancer cells and a syngeneic subcutaneous tumor model. Podophyllotoxin from Qinghai Sinopodophyllum hexandrum was identified by FT-UHR LC-MS. Its cytotoxicity against MFC gastric cancer cells was measured by CCK-8 assay. Cell proliferation, migration, invasion, and apoptosis were determined by colony formation, wound healing, Transwell, and flow cytometry assays, respectively. A syngeneic subcutaneous tumor model was established in 32 BALB/c mice for in vivo evaluation. Transcriptome sequencing was performed to profile gene expression in treated MFC cells, and Reverse transcription quantitative real-time PCR (RT-qPCR) verified key genes in MFC cells and tumor tissues. Podophyllotoxin significantly suppressed MFC cell proliferation, colony formation, migration, and invasion (P < 0.05), and induced early apoptosis (P < 0.05). In vivo, it inhibited tumor growth in a dose-dependent manner (P < 0.05). Transcriptome analysis showed that podophyllotoxin affected the DNA damage response pathway. RT-qPCR verified altered expression of DNA damage-related genes including Id3, Id4, Polq, Ddias, Gadd45a, Cip2a, Recql4, Rad54b, Mastl, and Fanci. Western blot confirmed downregulation of ID4 protein. Podophyllotoxin inhibits the growth of MFC gastric cancer cells and syngeneic subcutaneous tumors, probably by modulating the expression of DNA damage response-related genes. ID4 downregulation may serve as a potential target in this experimental model.
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