Inhibitory Effect Of Agkistrodon Acutus Antitumor Venom Component I For Human Oral Squamous Cell Carcinoma Cal27 Cells In Vitro And In Vivo

Objective: The objective of this study was to investigate the effect of tumor suppressing component I of Agkistrodon Acutus Venom on the proliferation,migration,invasion and apoptosis of oral squamous cell carcinoma(OSCC)Cal 27 cells,and the effect on tumor cell apoptosis and proliferation in vivo.Method:1.Cal27 cells were cultured in vitro.After 0.1mg/ml AAVC-I stimulation,the cell proliferation was determined by CCK8 method;the cell invasion was determined by Transwell chamber method;the cell migration was determined by cell scratch method;the apoptosis was determined by Annexin V-PI double staining method.2.Cal27 cells(1×107 cells/ml)were injected into the back of 12 male nude mice.After successful tumorigenesis,the experimental animals were randomly divided into AAVC-I group and control group.The experimental group was intraperitoneally injected with0.01ml/g normol saline diluted with 1.0mg/kg AAVC-I every other day,and the control group was injected with 0.01ml/g normal saline for 5 times.The volume of tumor was observed dynamically 48 hours after each administration and the dynamic growth chart was drawn.The mice were killed 48 hours after the last administration.The tumor tissue was stripped completely and weighed immediately to calculate the tumor inhibition rate.The tumor was sectioned and the cytology was observed by HE staining;the expression of Ki-67 antigen was observed by SP method;the expression of cleaved caspase 3protein was observed by immunohistochemistry.Results: 1.Compared with the control group,the proliferation of cal27 cells stimulated by AAVC-I was significantly inhibited,and when the time reached 96 h,the inhibition on the proliferation of tumor cells was stronger,which was time-dependent;the percentage of scratch healing was significantly reduced,and the migration ability of tumor cells was significantly reduced;the percentage of subventricular invasive cells decreased significantly;the percentage of apoptosis stimulated by AAVC-I was significantly increased.2.Compared with the control group,the growth rate of AAVC-I-treated nude mice was significantly inhibited,the tumor mass was smaller,the number of mitotic cells was significantly reduced under light microscope,the expression of Ki-67 antigen decreased,and the protein expression of cleaved caspase 3 was increased.Conclusion: AAVC-I can inhibit the growth of oral squamous cell carcinoma Cal27 cells,which is closely related to its inhibition of tumor cell proliferation and induction of tumor cell apoptosis.