Expression And Correlation Analysis Of USP9X, MCL-1 And Caspase3 In Cervical Squamous Cell Carcinoma

Objective: 1. To investigate the expressions and clinical significances of USP9 X, MCL-1 and Caspase3 in cervical lesions by immunohistochemistry. The relationships of the three factors in cervical squamous cell carcinoma(CSCC) were also analyzed in order to provide theoretic support for the study of the occurrence mechanisms of cervical carcinoma.2. To explore the correlations between the expression of USP9 X, MCL-1 and Caspase3 and clinicopathologic features of the CSCC. Changes of the expressions of the three in CSCC before and after the neoadjuvant radiation or chemotherapy were also analyzed in order to provide evidences for early diagnosis, clinical treatment and prognosis of CSCC.3. To observe the differerces of the expression of USP9 X, MCL-1 and Caspase3 in human cervical squamous carcinoma cell line Si Ha before and after treated with cis-platinum using cytological experimental methods. The effect of incereased sensitivity to cis-platinum in SiHa by USP9 X inhibitor WP1130 was also explored in order to provide a theoretical basis for seeking therapeutic target of CSCC.Method: 1. Collected 40 cases of NILM(who received total hysterectomy because of hysteromyoma), 40 cases of LSIL, 40 cases of HSIL, 85 cases of CSCC(45 cases of which were compared before and after radiotherapy and chemotherapy) from Jan. 2010 to Dec. 2014 in pathology department of the Affiliated Hospital of North Sichuan Medical College. The expressions of USP9 X, MCL-1 and Caspase3 in these tissues were detected by immunohistochemistry Envision method.2. The optimal concentration of cis-platinum and WP1130 effected on SiHa were screened by CCK8. The effects of different concentrations of cis-platinum, cis-platinum together with WP1130 on the inhibition rate of SiHa cells were studied. The effect of WP1130 on the expression of USP9 X in SiHa cells was detected by immunocytochemistry.3. The protein expressions of USP9 X, MCL-1 and Caspase3 before and after chemical drug in SiHa were detected by western blot and the correlations of the three were analyzed; The sensitivity of WP1130 to SiHa cells was also studied.4. The experiment data was analyzed through SPSS13.0 statistical software, P<0.05, having statistically significant difference.Result: 1. The rates of USP9 X and MCL-1 positive expression both increased gradually in NILM, LSIL, HSIL and CSCC which were closely associated with clinical stages of FIGO, pathological differentiation, depth of invasion, organ of CSCC and of Lymph node metastasis. The rates decreased after threpay(P<0.05). USP9 X and MCL-1 were positively correlated while the expression of Caspase3 had no obvious regularity.2. The optimal concentration of cis-platinum was 3ug/ml and WP1130 was 1.5μM, both treated 72 hours in SiHa. The survival rate decreased significantly in SiHa treated with cis-platinum and WP1130 compared with cis-platinum alone(P<0.05). Immunocytochemistry showed WP1130 could inhibit the expression of Usp9 x protein.3. The protein expressions of USP9 X, MCL-1 and proCaspase3 were significantly reduced in SiHa treated with 3ug/ml cis-platinum or(and)1.5μM WP1130(P<0.05); showing a positive correlation among the three. The protein level in SiHa which treated with cis-platinum combined with WP1130 was less than cis-platinum or WP1130 alone(P<0.05). WP1130 increased the sensitivity to cis-platinum in SiHa.Conclusion: 1. The expressions of USP9 X and MCL-1 were closely related to the occurrence and development of CSCC. The expressions of them were related to clinicopathologic factors of CSCC and could be affected by radiation or chemotherapy. The detection of the expressions of USP9 X and MCL-1 may be important markers for the diagnosis, assessment of therapeutic efficacy and prognosis of CSCC.2. The expression of MCL-1 was positively correlated with USP9 X.3. WP1130 sensitized CSCC to cis-platinum, providing a strong basis for the clinical integrated treatment of cervical carcinoma and suggesting that USP9 X may be a new target for the therapy of cervical carcinoma.