Inhibition Effects Of Pinellia Extract On Human Cervical Cancer Cell Lines And Associated Mechanisms

Cervical cancer is one of the most frequently gynecological neoplasma.With the prevalence of cervical HPV infection,the incidence of cervical cancer increased.More and more young women were suffered from cervical cancer.Cervical cancer is becoming one of severe diseases that could be harmful to women's health.Pinellia pedatisecta schott is a traditional Chinese drug in mainly regions of our country.It has pungent flavor,warm in nature and poisonous.Possess effects of dispelling wind and relieving convulsion,drying dampness to eliminate phlegm,drying dampness to eliminate phlegm,eliminating stagnation.It can be used to cure thanatophidia bite,nameless swelling and toxicum.In 1970's,Pinellia pedatisecta schott had been used to treat 247 cases of cervical cancer in our hospital,the effective rate was 81.53%.The exact pharmacological mechanism by which Pinellia pedatisecta schott offered in this treatment had not been identified.In this study,we have investigated the inhibition effect of Pinellia extract(PE) on the growth of cervical cancer cells and the associated mechanisms.There are three parts in the study:Partâ… Effects of PE on proliferation of the cervical cancer cell lines and mechanisms of action.Partâ…¡Apoptosis induction Effect of PE on the cell lines of cervical cancer and the associated mechanisms.Partâ…¢Effects of PE on the expression of P53 and HPVE6 in the cell lines of cervical cancer.Partâ… Effects of PE on proliferation of the cell lines of cervical cancer and mechanisms of actionObjective To observe the effects of PE on proliferation of cervical cancer cell lines CaSki and HeLa and its effect on the sensitivity of HeLa to DDP.Mechanisms of action were studied.Methods Cervical cancer cell lines CaSki and HeLa were incubated with different concentrations of PE for different time.We tested the effects of PE on cell proliferation by MTT assay.The effects of PE on morphology and ultrastructure were studied by phase-contrast microscope and electron microscope.PCNA was determined by immunocytochemistry experiment.In-cell Westem blot was used to estimate the relative protein amounts of phosphorylated ERK of CaSki and HeLa cervical cancer cells treated with different concentrations of PE for 15 minutes.Results PE could obviously inhibit the proliferation of CaSki and HeLa cells in a time and dose dependent manner.Low concentration of PE could elevate the inhibition effect of DDP on HeLa cells.Phase-contrast microscope and electron microscope revealed that the morphology and ultrastructure of CaSki and HeLa cells changed a lot.PCNA was down-regulated after treated with PE for 24 hours.PE could also inhibite phosphorylation of ERK in a dose dependent manner.Conclusions PE could suppress proliferation of cervical cancer cells and induce morphology and ultrastructure changes of CaSki and HeLa.PE could also elevate the sensitivity of HeLa cells to DDP.Inhibition effect of PE on PCNA and blocking of MAPK/ERK signal transduction pathway were involed in the mechanisms.Partâ…¡Apoptosis induction effect of PE on the cell lines of cervical cancer and the associated mechanismsObjective To investigate apopotosis induction effect of PE on the cell lines of cervical cancer and associated mechanisms.Methods The apoptosis and cell cycle of cervical cancer ceils were detected with flowcytometry(FCM) and DAPI stain.Bcl-2 protein was detected with immunofluorescence experiment.Results PE could induce cervical cancer cells apoptosis in a time dependent manner.The apoptosis rates of CaSki and HeLa cervical cancer cells were nearly 50%after treated with PE for 3 days,and up to more than 70%for 5days.But there was no change of cell cycle of cervical cancer cells after treated with PE.Apoptositic cells were also detected by DAPI. Bcl-2 protein was down-regulated after treated with PE for 24 hours.Conclusions PE could induce cervical cancer cells apoptosis.Bcl-2 was involed in the apoptosis induction process.Partâ…¢Effects of PE on the expression of P53 and HPV E6 in the cell lines of cervical cancerObjective To study the effects of PE on the mRNA and protein expression of P53 and HPV E6 in cervical cancer cell lines CaSki and HeLa.Methods RT-PCR and Realtime RT-PCR were used to estimate the relative amounts of P53 mRNA and HPV E6 mRNA.Western blot was used to measure P53 and HPV E6 protein levels before and after treatment with PE.Results After treated with 0.15mg/ml PE for 24hs,the expression of P53 mRNA increased significantly.The expression of HPV E6 mRNA and protein decreased after PE's exposure significantly.The expression of P53 protein increased also,but there was no statistical significance compared with the control group.Conclusion PE could up-regulate P53 gene's mRNA and protein expression,and could also down-regulate HPV E6 gene's mRNA and protein expression.The products of these genes could contribute to PE's effect on CaSki and HeLa,and then caused proliferation inhibition and apoptosis.