The Expression Significance And Correlation Of LC3B And CAIX During The Processes Of Autophagy In Cervical Squamous Cell Carcinoma

Research backgroundCervical cancer is one of the most common gynecological malignant solid tumors in the world, the incidence of which in china accounts for one of third of the worldwide incidence.In recent years, the incidence and mortality of cervical cancer peak age are dropping,and this change will seriously affect the reproductive health and life quality of young women. Persistent high-risk human papilloma virus(High risk-humanpapillomavirus, HR-HPV) infection is closely related to the incidence of cervical cancer.However, the incidence and development of cancer is generally the result of many biological factors working together, so HPV infection alone is not sufficient to lead to tumor development. Until now,the molecular mechanisms of Cervical cancer development is still not fully understood.Autophagy is a newly discovered mechanism of cell metabolism which have a complex connection with the development and treatment of tumors,which may play a different role in different tumor types, different parts of the same tumor, and different stages of the same type of tumor.Because Cervical squamous cell carcinoma of cervical cancer is the most common pathological type, which belongs to solid tumor,and hypoxia is one of its main physical characteristics.Hypoxia may reduce the sensitivity of cervical squamous cell carcinoma toradiotherapy and chemotherapy,but autophagy may enhance the sensitivity of tumor cells to anticancer chemotherapy and radiotherapy response.The relationship between the occurrence mechanism of autophagy and hypoxia in cervical cancer is unclear, and there are little related literature report, therefore, this study aims to detect the expression of autophagy marker protein LC3 B and hypoxia marker protein CAIX in normal cervical epithelial tissue and the cervical squamous cell carcinoma, and analyse the correlation and their relationship with clinical pathological characteristics of cervical squamous cell carcinoma,respectively. The preliminary discussion on the role of autophagy and hypoxia playing in the development of early cervical squamous cell carcinoma,and to analyse the influence of autophagy and hypoxia on the formation CD34 markers of microvascular density,are expected to give some new research reference value to explore the occurrence and development mechanism of the cervical squamous cell carcinoma.ObjectiveTo elucidate the expression of LC3 B and CAIX in normal cervical epithelial tissue and the cervical squamous cell carcinoma,and analyse the correlation and their relationship with clinical pathological characteristics of cervical squamous cell carcinoma, and to preliminarily discusse the role of autophagy and hypoxia playing in the development of early cervical squamous cell carcinoma,which are expected to provide a new target for the diagnosis and treatment of cervical squamous cell carcinoma.MethodUsing immunohistochemical method to detect the expression of LC3 B and CAIX in 72 cases of normal cervical epithelium and 126 example cervical squamous cell carcinoma tissue samples. semi-quantitative integral method is used to determine a positive result and using Western Blot to detect whether the results of LC3 B expression are consistent with immunohistochemical method. SPSS17.0 software is used for statistical analysis: using the Mann-whitney U test or Kruskal-wallis rank sum test analyze the relationship of LC3 B, CAIX expression with clinical pathological characteristics of cervical squamous cell carcinoma.The correlation analysis between the protein is used Spearman rank correlation test; the relationship between CAIX staining scores and LC3 B expression, and microvessel density of different groups of LC3 B and CAIX expression are all use analysis of variance. Using Graph Pad Prism 5.0 software to draw the relevant charts, and P <0.05 was considered statistically significant.ResultLC3B protein expressed mainly in middle and surface of normal cervical epithelial,and basal cell layer showed no staining. Its high expression rate was 70.8%(51/72),and the low and negative expression rate was 22.2%(16/72),7%(5/72)respectively; In cervical squamous cell carcinoma tissues,LC3 B protein high expression rate was 27%(34/126), and the low and negative expression rate was 43.6%(55/126),29.4%(37/126)respectively. The expression of LC3 B protein in cervical squamous cell carcinoma tissues was lower than that in normal cervical epithelium,statistically significant difference(P < 0.05).(2)CAIX protein expressed in a few basal cell layer of normal cervical epithelium, while intermediate and surface epithelium showed no expression. Its high expression was 8.3%( 6/72), 25% lower expression( 18/72), negative in 25%( 48/72); CAIX protein high expression in cervical squamous cell carcinoma was 29.4%( 37/126), low expression rate of 54%( 68/126), negative expression of 16.7%( 21/126),statistics showed : CAIX protein expression in cervical squamous cell carcinoma is higher than the normal cervical epithelium, and the difference was statistically significant(P <0.05).(3)The expression of LC3 B protein was positively correlated with CAIX protein and correlation coefficient is 0.403, P < 0.05.(4)MVD was mainly distributed in the edge of the tumor interstitial,with scattered distribution in minority cancer tissue.In the same serial tissue sections, high CAIX protein expression position was observed simultaneously with the high expression of LC3 B protein, MVD distributed in the cancer focus edge, while in the low expression of CAIX protein position, LC3 B showed low or no expression, and MVD located in the edge of foci and distrbute in lesions organization scatteredly. The mean of MVD in LC3 B protein different expression groups,such as in the negative, low, high LC3 B expression, the mean number were 31.92 ± 10.02,32.91 ± 9.99,31.06 ± 11.37,respectively. no significant difference between three groups(P>0.05). And the mean of MVD in CAIX protein negative expression, low, high expression groups were 26.90 ± 9.25,32.57 ± 9.91,34.24 ± 10.92,respectively. The three groups were statistically significant difference(P=0.028), SNK comparison between groups,the mean of MVD in CAIX low,high expression groups show no statistical difference, but there are differences between the high and negative expression group and between low andnegative expression.(5)LC3B protein expression was significantly correlated with tumor size(P = 0.001) and the degree of tumor differentiation(P = 0.035), so the difference was statistically significant. Hpwever, it is showed no significant correlation with age, FIGO stage, lymph node metastasis and preoperative Scc-Ag values; CAIX protein expression was only significantly correlated with tumor size,and there are statistically signifiConclusion(1)Autophagy with a lower incidence exist in cervical squamous cell carcinoma tissue,and the incidence of autophagy is only 27%.(2) The presence of autophagy in cervical squamous cell carcinoma, was positively correlated with the degree of tumor hypoxia.(3)The enhancement of autophagy in the cervical squamous cell carcinoma was associated with tumor size, and may not be associated with the formation of microvascular.(4) Hypoxia in Squamous cell carcinoma of the uterine cervix tissue was associated with the formation of microvascular,which promote the formation of the microvascular.