| 【Objection】1,To investigate the etiological role of HPV on the carcinogenesis of NSCLC and to study the relationship between HPV infection and clinical pathological characteristics.2,To study the relationship between HPV infection and the expression of Rb,CDK-2 protein, and to elucidate the possible molecular mechanisms that HPV infection induced carcinogenesis of NSCLC.3,To study the relationship between the expression of Rb,CDK-2 protein and carcinogenesis of NSCLC and clinical pathological characteristics.【Methods】60 cases of specimens were taken from Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. All patients didn't receive preoperative radiotherapy and chemotherapy. There were 29 cases of squamous cell carcinoma, 18 cases of adenocacinoma in 47 cases of NSCLC and high, medium and poorly differentiated lung cancer cases were 12, 18, 17; 28 cases of lymph node metastasis; 33 cases of smokers. There were 13 cases of benign pulmonary lesions (4 cases of pulmonary tuberculosis, 5 cases of inflammatory pseudotumor, 4 cases of bullous lung) as the control group. All specimens were selected to collect and record their age, sex, smoking history, lymph node metastasis and histological type and so on.1,The PCR methods were used to detect the HPV positive rate and the gene type by HPV16,18 type-specific primers in 47 cases of NSCLC and 13 cases of lung benign diseases.2,The immunohistochemical SP method was used to measure the expression of protein Rb and CDK-2 The protein expression levels of Rb,CDK-2 in 47 cases of NSCLC and 13 cases of lung benign diseases. Pictures for the result of IHC staining were gathered by system for study pictures. All data was dealt with SPSS for Windows 13.0 statistical software.【Results】1,The detection of HPV infection and protein Rb in 47 cases of lung cancer and 13 cases of benign pathological change lung tissues The PCR methods were used to detect the HPV positive rate and the gene type16/18. Detection rate of HPV DNA in lung cancer group was 42.55 %( 20/47) and that in benign disease group was 7.69 %( 1/13). There was a significant difference between the two groups (P<0.05).The immunohistochemical SP method were used to detect the expression of Rb protein and CDK2 protein。Negative expression rate of Rb protein was 68.09% (32/47) in lung cancer group, and there was significantly different with that in benign diseases 100% (13/13)(P<0.05), which suggested that the decrease of Rb protein might be one of the reasons for lung cancer. The expression of CDK-2 protein in lung cancer group was 82.98% ( 39/47), but there was no expression in the lung benign pathological change group. There was statistical difference between the two groups (P<0.05).2,The expression of protein Rb and CDK2 between HPV positive group and HPV negative group in 47 cases of lung cancerThe positive expression level of protein Rb in HPV positive lung cancers group was 45%(9/20). However, the positive expression rate of protein Rb in HPV negative of lung cancers was 85.19% (23/27). There was statistical difference between the two groups (P <0.05), from which we could imagine that HPV infection might directly or indirectly influence the expression and the effect of protein Rb.The expression level of protein CDK2 in HPV positive lung cancers group was 100%, and the expression rate of protein CDK2 in HPV negative of lung cancers was 70.37% (19/27). There was statistical difference between the two groups (P <0.05).3,The relationship among the infection of HPV and the expression of protein Rb and CDK2 with their clinic pathologic characteristics in 47 cases of lung cancer No correlation was found between HPV infection rate and age,sex,lymph node metastasis of patient. But there was a significant correlation among HPV infection rate with histological type,degree of differentiation,history of smoking(P<0.05). The HPV infection rate in squamous cell carcinoma 55.17% (16/29) was significantly higher than that in adenocarcinoma 22.22% (4/18). HPV infection rate in high,moderately and poorly differentiated NSCLC groups was 16.67%(2/12),66.67% (12/18) and 35.29%( 6/17). There was statistical difference among the three groups (P <0.05) and it was higher in moderately and poorly differentiated NSCLC groups. HPV infection rate in smoking lung cancer patients'was 54.54%( 18/33) which was significantly higher than that in non-smokers 14.29% (2/14).IHC methods were used to detect the expression of Rb protein and CDK2 protein in 47cases of NSCLC tissues and analysis of their relationship with clinical pathological parameters (age, sex, smoking history, histological type, differentiation degree, lymph node metastasis). There was no relationship between Rb negative expression and the clinical pathological characteristics of NSCLC. The relationship was found between the expression of CDK-2 protein and degree of carcinoma differentiation. Expression rate of CDK-2 protein in high differentiation lung cancer was 58.33% (7/12) and 83.33% (15/18) in middle differentiation lung cancer, which was 100% (17/17) in low differentiation lung cancer. It prompted that there was a relationship between the expression of protein CDK2 and the differentiation of the lung cancer (p<0.05). But there was no relationship between the expression of CDK-2 protein and the other clinical pathological characteristics.【Conclusion】1. HPV16/18 high-risk type infection of NSCLC tissue was significantly higher than benign pathological change lung tissues, and HPV infection was related to histological type,degree of carcinoma differentiation and history of smoking. It suggested that HPV infection was one of important etiological factors in carcinogenesis of NSCLC, especially squamous cell carcinoma. 2. The expression of Rb protein was significantly different between NSCLC tissues and benign pathological change lung tissues, and in the lung cancer group the abnormal expression of Rb protein associated with HPV infection, there was a relationship between aberrant expression of Rb in lung cancer group and HPV infection. HPV might form a complex with Rb protein namely E7-PRb compound, and then made PRb degradation and inactivation, consequently HPV participate in progression of NSCLC.3. The expression of CDK2 protein was significantly different between NSCLC tissues and benign pathological change lung tissues, and there was a relationship between high expression of CDK-2 protein in lung cancer group and HPV infection. This suggested that HPV might act on cell cycle regulators and induce carcinogenesis of NSCLC by breaking the balance the cell cycle.4. There was relationship between expression of Rb,CDK-2 protein and NSCLC with HPV positive. They might have a cooperation role in carcinogenesis of NSCLC. |
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