Expression And Clinical Significance Of CLC-3 MRNA In Cervical Cancer

BackgroundCervical cancer is one of the most common cancers affecting women world-wide, it remains a leading cause of cancer-related death for women in developing countries. HPV infection plays a necessary role in cervical carcinogenesis while viral infection alone may not be sufficient for the development of cervical cancer. There must be some other cofactors have an influence upon the development of cervical carcinoma.Chloride ion channel is the major anion in the body and has close relationship with the proliferation of cell. It has massive expressions in many tumor cells. Chloride channels are classified based on structural differences. CLC-3 is one kind of chloride channel (voltage-gated chloride channels), CLCs are generally gated by transmembrane voltage, but also show gating in response to pH, cell swelling, or phosphorylation. Currently, CLC-3 chloride channel express higher in S phase than in GO phase during Human cervical cancer SiHa cell's cycle progression. While there have no clear that CLC-3mRNA express whether or not in vivo cervix. The aim of this thesis was to detect the expression of CLC-3 gene in order to find their relation with cervical cancer and the prognosis by RT- PCR.PurposeTo study the expression and clinical significance of CLC-3mRNA in cervical cancer.MethodsTotal RNA samples used in this study were isolated from 60 patients with invasive cervical cancer who underwent biopsy or curative resection, and another 10 pairs cervical cancer specimens from the same patients who before and after radiochemotherapy two weeks at the First Affiliated Hospital of China Medical University. Besides, we collected 10 cases of normal cervical tissue. These tissue samples were obtained after hysterectomy because of benign uterine myoma.Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of CLC-3mRNA in 60 patients with cervical cancer, 10 patients with cervical cancer before and after radiochemotherapy , and 10 patients with normal cervical tissue. Total RNA extraction, cDNA and subsequent PCR amplifications according to the RNA PCR Kit(AMV)Ver3.0 instruction.PCR products (5ul) were electrophoresed with 2% agarose-1×TAE gels containing 0.5 ug/ml ethidium bromide and scale gray level for data analysis and statistics. Paired t-tests or nonparametric tests were used to evaluate statistical significance. Graphs and further analysis were generated with SigmaPlot5 software (Aurora,CO). Differences were considered as significance when P values were less than 0.05(P<0.05).Results1,The ratio of gray level of CLC-3mRNA were significant different between the normal cervical tissues and cervical cancers, (P=0.001).2,The ratio of gray level of CLC-3mRNA in the poor differentiation cancer was higher than that of the middle and high differentiation cancer (P=0.002). Same as tissue differentiation, the ratio of gray level of CLC-3mRNA was higher in stages I～II than that in stages III～IV, there were significant differences between them (P=0.001). But there was no distinction among age groups (P=0.665).3,CLC-3mRNA was higher before radiotherapy and chemotherapy than that after radiochemotherapy, they have significant differences(P=0.003).Conclusion1,CLC-3mRNA participates in the proliferation of cervical cancer cell, and plays an important role in tumorgenesis of cervical cancer.2,CLC-3mRNA have a positive correlation with clinical stages and pathological grades of cervical cancer.3,Radiochemotherapy inhibits the expression of CLC-3mRNA in cervical cancer .