Experimental Study Of Vitro Effect To Cervical Carcinoma Hela Cells By Proteasome Inhibitor Lactacystin

Objective: This article aims to research the vitro effect to cervical cancer Hela cell by proteasome inhibitor Lactacystin,and to explore its possible mechanism, also to provide new theoretical ideas and experimental evidence for the treatment of cervical cancer.Methods: MTT and Hoechst 33342 staining to detect the proteasome inhibitor Lactacystin for cervical cancer Hela cells'growth,proliferation and apoptosis; and detect cell injury-related signals and gene expression changes by means of Western Blot and RT-PCR.Results:(1) MTT results showed that: compared with the control group, in LAC 1.25μM group (86.5±0.09 (%)), LAC 2.50μM group (65.4±0.11 (%)), LAC 5.00μM group (52.3±0.15 (% )), LAC 10.00μM group (40.2±0.12 (%)) and LAC 20.00μM group (22.9±0.13 (%)), the proliferation rate of Hela cells was significantly decreased( (P <0.05).(2) Hoechst 33342 staining indicated that: With the concentration of LAC increased, the phenomenon of chromatin condensation,nuclear shrinkage and nuclear fission was more obvious,but in control group,the nuclear of Hela cells kept integrity and the chromatin of Hela cells distributed uniformly.(3) Western Blot test results showed that: contrast to the control group, with the concentration of LAC increased, the expression of Bcl-2 protein decreased gradually, and the expression of Bax protein increased rapidly, in 10.00μM concentrations,the ratio of Bax/Bcl-2 protein significantly increased (P <0.05), while, with the concentration of LAC increased, the expression of Caspase-3 protein also increased visible (P <0.05).(4) RT-PCR electrophoresis results showed that: compared with the control group, with the concentration of LAC increased, Caspase-3 mRNA expression was significantly increased, the expression of Bcl-2 mRNA decreased rapidly, while Bax mRNA expression was significantly increased,and the ratio of Bax / Bcl-2 mRNA increased more markedly (P <0.05).Conclusion: (1) In vitro experiments, the proteasome inhibitor Lactacystin inhibits the proliferation of cervical cancer Hela cells.(2) In vitro experiments, the proteasome inhibitor Lactacystin induces apoptosis in cervical cancer Hela cells.(3) The proteasome inhibitor Lactacystin maybe induce apoptosis in cervical cancer Hela cells mainly through the mitochondrial pathway,and maybe through up-regulating the expression of Caspase-3 and the ratio of Bax/bcl-2.(4) Inhibition the function of proteasome,can inhibit the proliferation of cervical cancer Hela cells significantly and induce apoptosis.In short, the vitro experiments confirmed that the proteasome inhibitor Lactacystin can inhibit the proliferation of cervical cancer Hela cells and induce apoptosis, the machanism of apoptosis may be mainly through the mitochondrial approach, with up-regulating the expression of Caspase-3 and the ratio of Bax/bcl-2.