AHVAC- inhibits the proliferation in human primary gastric cancer cells / 中国临床药理学与治疗学

ABSTRACT

AIM: To explore the effect of inhibiting proliferation and inducing apoptosis in human primary gastric tumor by the Agkis-trodon halys venom anti-tumor component (AHVAC-). METHODS: Human primary gastric cancer cells were isolated by trypsin digestion, serum-free culture, and purified by differential adherence method, and cells were identified by immunohistochemistry. Cell proliferation and toxicity assay (CCK-8) was used to detect the inhibition rate of AHVAC- in different concentrations of primary gastric cancer cells. Immunohistochemistry was used to verify the apoptosis of human primary gastric cancer cells induced by AHVAC- and the morphological changes were observed by Hematoxylin-Eosin staining (HE staining). AHVAC--induced primary gastric cancer cell transformation rate was detected by flow cytometry Annexin V/PI double staining.RESULTS: Seven human primary gastric cancer cells were successfully isolated and purified, and 11 cases failed. Immunohistochemical identifications of carcinoembryonic antigen (CEA) and broad-spectrum keratin protein (AE1/AE3) were positive for both antibodies. AHVAC- inhibited the proliferation of human primary gastric cancer cells and showed a dose-dependent effect (P<0.01). Immunohistochemistry showed that the expression level of cysteine aspartic protease-3 (Caspase-3) up-regulated with the increase of AHVAC- concentration. HE staining showed that with the increase of AHVAC- concentration, the cell gap increased, nuclear pyknosis, and apoptosis cells increased. Flow cytometry showed that the apoptosis rate of human primary gastric cancer cells up-regulated with the increase of AHVAC- concentration (P<0.05). CONCLUSION: AHVAC- can inhibit the proliferation and induce apoptosis in human primary gastric cancer cells in a dose-dependent manner.