Tianjin Medical Journal

Tianjin Medical Journal ›› 2019, Vol. 47 ›› Issue (5): 454-458.doi: 10.11958/20182153

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The molecular mechanism of PDCD4 knockdown facilitating cisplatin resistance in gastric cancer through pAKT/pGSK3β pathway

LIU Dan1, TANG Zhi-ming1, WU Fu-yun1, ZHAO Hong-yan1, KE Jing1, XIANG Peng3, JIN Jia-qing4, LI Shan1, 2△   

  1. 1 Institute of Basic Medical Sciences, Hubei University of Medicine, Shiyan 442000, China; 2 Department of Integrated Medicine, General Hospital of Dongfeng, Hubei University of Medicine; 3 Department of Orthopedics, People’s Hospital of Sheyang County; 4 Department of Clinical Laboratory, Wuhan Eighth Hospital △Corresponding Author E-mail: lishanhbmu@126.com
  • Received:2018-12-29 Revised:2019-03-15 Published:2019-05-15 Online:2019-05-15

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Abstract: Abstract: Objective To investigate the role of programmed apoptotic factor 4 (PDCD4) deficiency in the cisplatin induced apoptosis, and provide the new target marker for cisplatin resistance in gastric cancer. Methods ShNC/shPDCD4 vector was stably transfected into human gastric cancer cell line SGC7901 with or without cisplatin. The cells were divided into shNC group, shPDCD4 group, shNC with cisplatin group, shPDCD4 with cisplatin group for the following experiments. Real-time PCR and Western blot assay were used to detect the mRNA and protein expression levels of PDCD4. The caspase-3 activity kit and Hoechest dying with immunofluorescence were used to measure the cell apoptosis in vitro. The protein levels of pAKT and p-GSK3β were detected by Western blot assay. After combined with Akt inhibitor, the cells were divided into shNC with cisplatin group, shPDCD4 with cisplatin group, LY294002 inhibitor group and Wortmannin inhibitor group, and the protein levels of PARP were detected by Western blot assay. Results The results of qRT-PCR and Western blot assay confirmed that the mRNA and protein expressions of PDCD4 decreased significantly in SGC7901 cells, which stably transfected with ShPDCD4 plasmid, decreased significantly. The SGC7901 gastric cancer cell line that stably transfected with ShPDCD4 was successfully constructed in vitro. Caspase-3 activity and apoptosis were significantly decreased in shPDCD4 with cisplatin group than those of shNC with cisplatin group (P＜0.05). The expressions of pAKT and pGSK3β increased due to the deficiency of PDCD4 expression (P＜0.05). After blocking the signal pathway with Akt inhibitors (LY294002 and Wortmannin), the relative expression level of PARP was up-regulated (P＜0.05). Conclusion The down-regulation of PDCD4 expression can reduce the apoptosis of gastric cancer cells induced by cisplatin through pAKT/pGSK3 beta pathway, thus promoting the formation of cisplatin resistance.

Key words: cisplatin, drug resistance, neoplasm, stomach neoplasms, programmed apoptosis factor 4, protein kinase B, glycogen synthase kinase 3β