Regulatory Effect Of MicroRNA-221 On ASPP2 Protein In Cervical Cancer

Expression level of ASPP2 and mi RNA that possibly target it in cervical carcinomaObject: To detect the expression of ASPP2 and target miRNA in cervical carcinoma and uterine leiomyoma.Method: The expression of ASPP2 in m RNA level was detected in fifty-five cases of cervical carcinoma tissues and twenty-three cases of uterine leiomyomas as control by using RT-PCR method. Bioinformatics software was used to predict the target mi RNA of ASPP2, and the expression of the target mi RNA in m RNA level was detected in cancer tissues.Result: The expression of ASPP2 in mRNA level was obviously lower in cervical cancer tissue than that in uterine leiomyoma, and the difference is statistically significant(P < 0.01). In addition, with the combination of clinical data, the expression of ASPP2 m RNA was higher in stage I of cervical cancer tissues than that of stage II and III, and the difference is statistically significant(P=0.0361). Through the application of biological information software, mi RNA-370, mi RNA-221, mi RNA-222 and mi RNA-205 were predicted to be the possible target mi RNA of ASPP2. The result of testing the expression of mi RNA-221 in cervical cancer tissues showed that mi RNA-221 m RNA in cervical cancer tissue was higher than that in control group, and the difference was statistically significant(P=0.026).Conclusion: The expression level of ASPP2 in cervical cancer was lower than that in normal tissues, and the level would decreased with the deterioration of the diseased lesions, suggesting that ASPP2 may play an important role in the occurrence and development of cervical cancer. As the expression of mirna-221 serving as the possible ASPP2 targeted mi RNA was increased in cervical cancer, it is possible to participate in the regulation of cervical cancer.Part II Validation of mi RNA-221 in the regulation of ASPP2 in cervical cancerObject: To construct the ASPP2 3 'UTR luciferase reporter gene plasmid, and to verify the relationship between mi RNA-221 and ASPP2 in cervical cancer.Method: The constructed ASPP2 3 'UTR luciferase reporter gene plasmid and mi RNA-221 mimics was transfected into cervical cancer cells Siha, and the fluorescence intensity was detected by luciferase reporter assay kit.Result: The construction of ASPP2 3 'UTR luciferase reporter gene plasmid was successful, and the result of reporter gene showed that the fluorescence intensity decreased by nearly 40% after the transfection of mi RNA-221 mimicsConclusion: MiRNA-221 can be directly combined with ASPP2 3 'UTR to regulate ASPP2.Part III To study the function of ASPP2 in cervical cancer and the regulation effect of mi RNA-221 on ASPP2Object: To study the cellular function of ASPP2 in cervical cancer cells, and to study the changes of cell function under the control of mi RNA-221.Method: The ASPP2 over-expressed vector TP53Bp2 was constructed, and then transfect into cervical cancer cell lines Siha. A series of cellular functional experimental methods including CCK8, cell scratch test, Transwell assay and so on were used for studying the function of ASPP2 on cancer cells. Micro RNA-221 mimics and ASPP2 expressed vector TP53Bp2 were transfected into cervical carcinoma cell line Si Ha together, after the regulatory effect of micro RNA-221 on ASPP2 protein was explored by cellular functional experiments including CCK8, cell scratch assay, Transwell assay and so on.Result: Since ASPP2 over-expressed, the amount of apoptotic cervical cancer cells Siha was increased and the invasiveness of the cells is decreased. After transfection of Micro RNA-221-3p into the cervical cancer cells over-expressed ASPP2, the apoptosis of these cancer cells is lower than that of untreated ASPP2 over-expressed Si Ha cell lines. Further, the invasiveness of ASPP2 over-expressed cervical cancer cells with the transfection Micro RNA-221-3p is greatly improved compared to the ASPP2 overexpressed Si Ha cell lines without any treatment.Conclusion: ASPP2 in cervical cancer has the role of inhibiting cancer, can inhibit the proliferation of cancer cells, reduce the invasion of cancer cells, and the migration of cancer cells is not obvious, pending further study. Under the control of ASPP2 to mi RNA-221, the inhibition effect of ASPP2 in cervical cancer can be reduced, the relative increase of cancer cell proliferation, and increased the invasion of cancer cells.