Effect Of Biological Behavior And Clinical Significance Of Maspin Gene On Cervical Squamous Carcinoma SiHa Cell

Objective:1. Construct the recombinant eukaryotic expression vector of maspin and stably transfected it into cervical carcinoma SiHa cell lines.2. To detect the expression changes of maspin in SiHa cells.3. To evaluate the effect of mammary serine protease inhibitor (maspin) overexpression on human cervical squamous carcinoma (SCC) SiHa cell proliferation and apoptosis in vitro.Methods:Recombinant plasmid pcDNA3-maspin was stably transfected into human cervical SCC SiHa cell. Maspin mRNA was determined by real time RT-PCR and RT-PCR whereas maspin protein was detected by Western blot analysis and SABC immunocytochemistry (IC). Cell proliferation activity was measured by MTT method. Apoptosis rate and cell cycle distribution were detected by flow cytometry to understand the changes in the cell biological characteristics.Results:The strengthened expression of the maspin gene in the SiHa cell was confirmed by real time RT-PCR, RT-PCR, Western blot, and IC (P<0.05). Suppressed proliferation activity and increased apoptosis rate of SiHa-m (maspin stable transfected) versus SiHa and SiHa-vector cell (SiHa-pc3) were shown by MTT and flow cytometry (P<0.05). SiHa and SiHa-pc3 had no statistical significance (P>0.05).Conclusions:The results showed that maspin gene can significantly inhibit human cervical SCC SiHa cell proliferation and effectively slow cancer growth. Maspin may be a new molecular target in the gene therapy of human cervical SCC. However, the mechanism of maspin in inhibiting tumor cell growth and regulating apoptosis needs further discussion.