The Effect Of MiR-224 On The Growth Of Cervical Cancer Xenografts In Nude Mice By Affecting Autophagy

Objective:To investigate the effect of miR-224 on the growth of SiHa and C33 A cervical cancer xenografts in nude mice.Methods:(1)The expression of miR-224 was over express or down regulation through transfection.Then it was detected by Flow Cytometry and Quantitative Real-Time Polymerase Chain Reaction(qRT-PCR).The every cell lines were divided into five groups:miR-224 mimics group,miR-224 mimics negative control group,miR-224 inhibitor group,miR-224 inhibitor negative control group and negative control group.(2)The above 10 groups of cervical cancer cells were prepared as cell suspension with phosphate buffer saline and mixed with matrigel.The mixed cell suspension was injected subcutaneously into the back of the neck of the nude mice by insulin needle.The life activity of tumor bearing mice was observed and the changes of tumor volume were estimated.The differences of the transplanted tumor size in every group were compared by the tumor growth curve and the quality of transplanted tumor at the end of observation.(3)Western-blot and qRT-PCR were used to detect the expression of p62 and LC3 in the tumor.Results:(1)The expression of miR-224 in the miR-224 mimic group of SiHa and C33 A cellswas significantly higher than that in the negative control group(p < 0.05);The expression of miR-224 in the miR-224 inhibitor group was significantly lower than that in the negative control group(p < 0.05).(2)At the same time point in the intervention 5 group,the tumor volume of SiHa cells was larger than that of C33 A cells(p < 0.05).SiHa and C33 A cell transplantation tumor models were compared among five groups respectively.At the first measurement(the 8th day),there were significant differences between the experimental group and the control group,that is,the tumor volume of miR-224 mimic group was larger than that of the negative control group(p < 0.05),while the tumor volume of miR-224 inhibitor group was smaller than that of the negative control group(p < 0.05),and the significant differences persisted.At the end of the observation,the quality of SiHa cell transplanted tumor was higher than C33 A cell transplanted tumor(p < 0.05).SiHa and c33 a cell models were compared among five groups.The quality of transplanted tumor in mir-224 mimic group was significantly higher than that in control group(p < 0.05),and that in miR-224 inhibitor group was significantly lower than that in control group(p< 0.05).(3)The results of Western-blot and qRT-PCR showed that the expression of p62 in miR-224 mimic group was higher than that in the control group,and the expression of LC3 II in miR-224 inhibitor group was lower than that in the control group,and the expression of LC3 II in miR-224 inhibitor group was higher than that in the control group(p < 0.05).Conclusion:miR-224 promotes the growth of cervical cancer xenografts in nude mice by inhibiting autophagy.