| Cancer is a serious threat to human life and safety of one of the diseases.To seek efficient and low toxicity of anticancer drugs and treatment of the scientific community has been on the hot spots. Scorpion venom is a biological toxin, because of its wide range of physiological effects closely watched. This paper aims to study the destruction about scorpion venom on cultured human liver cancer cells (SMMC 7721) and the Hela cell ,and their mechanisms were discussed for the development of anticancer drugs and scorpion toxin of anti-tumor clinical applications experimental basis.Objective: detected using scorpion venom poisoning and the rough part of the purification of human liver cancer cells (SMMC 7721) and Hela cell growth inhibition;to detect apoptosis-related proteins of human liver cancer cells (SMMC 7721) and Hela cells, to explore the scorpion venom for the mechanism,verify the sensitivity of the scorpion venom to the two kinds' cell lines.Methods: Human liver cancer cells (SMMC 7721) and Hela cell line were given by scorpion venom and purification in accordance with the concentration gradients of 24 hours, and then were detected the survival rate by MTT. Detect apoptosis-related proteins' expressed levels of caspase-3 and bcl-2 proteins with western blotting and immunocytochemistry method .And Actin was as internal parameter. verify the sensitivity of the scorpion venom to the two kinds' cell lines with the immunofluorescence methods.Results: 1. MTT results:(1.1) As the toxic effects of scorpion venom through 24 hours, the survival rate of crude drug group of liver cancer cells (SMMC 7721) was lower than the control group significantly (P<0.05), and as the concentration of scorpion venom increased (0.28,0.70,1.4, 2.8 and 5.6μg/ml), the extension of time (4h, 8h, 12h, 24h),liver cancer cell survival rate decreased gradually ,with a gradual change.(1.2) As the part of purification of scorpion venom through 24 hours, the survival rate of purified human liver cancer cells group (SMMC 7721) were lower than the control group significantly (P<0.05), and with the increasing concentration of the part of purification of scorpion venom (0.28,0.70,1.4,2.8and5.6μg/ml), the extension of time (4h,8h,12h,24h),liver cancer cell survival rate decreased gradually ,with a gradual change.(1.3) The effect of the purification of scorpion venom was weaker than the crude group.(2.1) As the toxic effects of scorpion venom through 24 hours,Hela cells of crude drug group had no significant differences with the control groups with the concentration of 0.28,0.70,1.4μg/ml;had differences with the control groups (P<0.05) with the concentration of 2.8and5.6μg/ml.With the role of extended time (4h,8h,12h,24h), survival rate of Hela cells decreased gradually ,with a gradual change after 8h.(2.2)As part purification product of scrpion venom(0.28,0.70,1.4,2.8 and 5.6μg/ml),the survival rate of Hela cells had no significant difference with the control group whihin 24h.(2.3)Hela cells had less sensitivity than the human liver cancer cells (SMMC 7721) to scorpion venom.2.Western blotting results:After treated by scrpion venom with the concentration of 2.8μg/ml, apoptosis protein caspase-3 increased and anti-apoptosis protein bcl-2 decreased in human liver cancer cells group (SMMC 7721).3.immunocytochemistry results:After treated by scorpion venom with the concentration of 2.8μg/ml within 8h,apoptosis protein caspase-3 increased and anti-apoptosis protein bcl-2 decreased in Hela cells. 4 immunofluorescence resultsAfter treated human liver cancer cells (SMMC 7721) and Hela cells 8h with the crude scorpion venom that were handle with the fluorescent marker.human liver cancer cells (SMMC 7721) were with more fluorescent material than Hela cells under fluorescent microscope.Conclusion:1.SVC and some rough purification component inhibited the growth signficantly 0f human liver cancer cells (SMMC 7721) . With the increasing concentration of scorpion venom(0.28,0.70,1.4,2.8and5.6μg/ml) and the extension of time(4h,8h,12h,24h),the survival rate decreased gradually within 24h.Although the part of purificatied scrpion venom had a single molecular weight relatively,but it had less weaker role than the crude drug group.2. When the concentration of scorpion venom reached 2.8 and 5.6μg/ml,the processing time was more than 8h,Hela cell survival rate fell gradually,but it was significantly lower than the sensitivity of human liver cancer cell(SMMS 7721).3. The scorpion venom can inhibit the cell growth of SMMC7721 and Hela cells through the machanism of apoptosis in preliminary view.
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