The Influence Of Cervical Cancer HELA Cell With Electrochemical Therapy

The influence of cervical cancer HELA cell with electrochemical therapyObjective:Through vitro experiment,we observed and analyze ion channels,cell growth,apoptosis and their relationship when electrochemical treatment perform on cervical carcinoma cell line,to further explore the treatment of cervical cancer electrochemical therapy mechanism.Methods:Cell Count growth curve to the observation(5V,10C) Electrochemical Treatment dose role,cathode,anode and(5V,10C)after electrochemical treatment and added to the K + channel opener on the cathode,anode cells,not treated with cervical cancer cell growth trend changes.MTT assay(MTT)to compare the above cell growth inhibition; Whole-cell patch-clamp mode measured by(5V.10C)electrochemical treatment of the anode and cathode cells with untreated cervical cancer cells,K + current changes.RT-PCR(RT-PCR)was measured by(5V,10C), (10V,10C),(10V,5C),(5V,5C)Electrochemical Treatment dose role,Anode and cathode cells not treated with cervical cancer cells,compared to its potassium ion channel gene KCNJ16.calcium channel protein gene CACNA1G.and the chloride channel gene expression CLCN3mRNA; Semi-quantitative PCR(electrophoresis gel image analysis)were measured to calculate the dose electrochemical cell cathode and anode cell KCN J16,CACNA1G,CLCN3mRNA the relative expression of values,and compare the difference.Results:(1)to control the time factors, (5V,10C)electrochemical treatment group cathode and anode group, (5V,10C)electrochemical treatment and add the K + channel opener on the cathode and anode Group Group and the control group cell count comparison between the results of a significant statistical difference(P＜0.01) by the growth curve showed that the control group with the time change cells gradually increased;(5V.10C)after treatment cathode and anode group Cells with time change has decreased,with the Other three groups are clearly different;(5V.10C)electrochemical treatment and add the K + channel opener of the Group cathode and anode cell few first decline After the third day of a gradual increase in the final group with normal cell growth trend.(2)in cervical cancer Hela cells,using the whole-cell patch-clamp recording of sustained activation of outward potassium currents.Current such activation faster deactivation very slow,with outward rectifier characteristics,L.approximately Department.began around can be identified potassium currents,with instructions voltage step up,gradually increase the opening performance of the outward current values increase,hence the voltage-dependent.By (5V,10C)Electrochemical Treatment dose after the cathode and anode and the control group cells,Cell outward rectifier potassium channel was inhibited,the control voltage 10mv,20mv,30mv,40mv,50mv,the experimental group compared with the control group,the K + channel intensity had dropped,from the I-V curves showed that Compared with the control group,by electrochemical treatment,the cathode and anode Group I-V curves downward.(3)Semi-quantitative PCR results show.:the control group and by electrochemical treatment(5V,10C)cathode and anode, (10V,10C)anode KCNJ16 relative value compared to treatment KCNJ16 expression,The difference was statistically significant(P＜0.05)was statistically significant;drop:(5V,10C)anode "(10V,10C)cathode and anode,the difference was significant(P＜0.05)was statistically significant,After the differences between the two no significant difference(p＞0.05),the control group and the other groups with no significant difference;Compared with the control group,(5V,10C) electrochemical treatment,the Department CACNA1G cathode increased relative expression,the difference was statistically significant(P＜0.05),the group I was no significant difference;(5C,10V)after treatment cathode,(10C,10V)cathode cells after treatment group and the control group and the rest of the group cells,CLCN3 expression increased, the difference was significant(P＜0.05)which this group is the difference between statistical significance(P＜0.05),the gene expression in the former than the latter increased significantly.I of the group and the control group compared the gene expression of no significant difference.Conclusion:(1),electrochemical treatment of a certain dose intensity can raise KCNJ16,CNCL3.CACNA1G change,in which the KCNJ16 for the inward rectifier potassium channel coding genes, change occurred mainly in cathode,and electrochemical mechanism related to the treatment;CACNA1G and CNCL3 of T-type calcium,channel gene encoding and capacity control chloride channel-coding because of the cathode may have increased the expression,and electrochemical treatment of the mechanism needs further study.(2)Whole-cell patch clamp recording revealed outward rectifier K~+currents in the electrochemical treatmentto be restrained..And it is one of a antitumor mechanisms of electrochemical therapy.