Relationship of PCNA, COX-2 and Microvessel Density in Cervical Squamous Cell Carcinoma
|Course||Obstetrics and Gynaecology|
|Keywords||Cervical neoplasm CD105 proliferation cell nuclear antigen (PCNA) cyclooxygenase-2(COX-2) microvascular density (MVD)|
Background and objectiveCervical cancer is the common woman malignant tumor second only breast cancer in the world. Age of patients is double humps, 35～39y and 60～64y. According to the estimation of international cancer center, neopathy of cervical cancer account 9.8% in all tumors and it is a tendency that the proportion of young patients is rising. Cervical cancer has long ante-cancer stage, but etiological factor is not clear. Domestic and abroad studies indicate it relates to early sex behavior, chaos sex behavior, early delivery, copiousness, HPV infection and economic status.Tumor angiogenesis theory considers growth and metastasis of tumor depend on tumor angiogenesis and chemical signals produced by tumor metabolism activate vascular endothelial cell to promote tumor angiogenesis. Entity tumor only can grow to 1~3 mm and can’t metastasis without angiogensis. Cervical caner’s main route of metastasis is creep what very depend on angiogenesis. Recently studies dedicate carcinogenesis and developments of cervical cancer relate to tumor angiogenesis closely which can be used to judge prognosis and observe curative effects.Microvessel density (MVD) is considered to be the most effective objective marker of tumor angiogenesis. CD105 (Endoglin) is a new specific marker of microvessel which only mark new vascular endothelial cells. PCNA is a nuclear protein used to detect proliferating cell which expression has a distinct difference in multiplication cycle. Cyclooxygenase-2 is a significant rate-limiting enzyme in breakdown arachidoncic acid to every endogenous prostaglandin (PGs) which can cause inflammatory reaction and promote tumor angiogenesis. In this study, we probe the relationship between tumor angiogenesis and cell proliferating in carcinogenesis and development of cervical cancer through detecting CD105, PCNA and COX-2 in normal cervical tissues, precancerous changed tissues and cervical squamous cell carcinoma tissues by immunohistochemical SP method to provide evidence to choice reasonable modus operandi and judge prognosis in clinic.Materials and methods1 Materials: cervical specimens were obtained from 96 patients including 32 normal cervical tissues, 7 cervical intraepithelial neoplasia (CIN) I, 9 CIN II, 18 CINIII and 30 cervical squamous cell carcinomas. All patients did not accept radiotherapy or chemotherapy before operation and all specimens were confirmed by pathology. Age is cervical squamous cell carcinoma patients from 24～65 years (mean 43 years). Clinical stage using International Federation of Gynecology and Obstetrics (FIGO) 2000 clinical stage standard contains 7 I A,11 I B, 7II A, 5IIB. 26 were under pelvis lymph node excision and 5had lymph node metastasis in 30 cervical cancer.2 Methods: immunohistochemical SP method was used to detect expression of CD105, PCNA and COX-2 in normal cervical tissues, CIN tissues and cervical squamous cell carcinoma tissues. Counting microvessl marked by CD105 to calculate MVD and using PCNA masculine cell to calculate proliferation cell index (PI). Calculate COX-2 gray degree by special image analysis system. All data was analyzed by SPSS 11.0 package. Significance level is a=0.05.Results1 The positive expression of CD105 was mainly localized cytomembrane or cytoplasm of vascular endothelial cells in the interstitial tissue around the tumor nests.MVD marked by CD105 is 5.13±2.30 in normal cervical tissue, 11.57±1.40 in CINI,11.44±3.54 in CIN II, 14.50±2.28 in CINIII and 15.40±2.90 in cervical cancer. In cervical cancer, MVD marked by CD105 in well differentiated is 14.71±2.43; 13.77±2.24 in moderately differentiated and 18.00+2.16 in poorly differentiated cervical cancer and the difference between poorly differentiated type and other two types has statistical significance. According to FIGO 2000 standard it is 14.17±2.68 in I and 17.25±2.18 in II and the difference between them has statistical significance. MVD is 18.00±2.35 in lymph node metastasis and 15.10±2.76 in no lymph node metastasis and the difference between them has statistical significance. Dividing patients into two groups by age whether over 40 and MVD in young group is 16.64±2.20 contrasting to 14.68±3.06 in old group and the difference between them has no statistical significance too. 2 PCNA mainly express in cell nuclear in period of proliferation which was dyed to yellow. It can be seen in tumor cell nuclear, atypical hyperplasia cell nuclear and some normal cervical squamous epithelium cell nuclear in basal lamina in period of proliferation. PCNA express diffusely in cancer nest displaying masculine cell lumping distribution.PI is 4.97±2.65 in normal cervical tissues, 14.14±3.02 in CINI,16.78±2.33 in CINII, 31.22±3.61 in CINIII and 44.27±3.74 in cervical cancer. In cervical cancer, PI in well differentiated is 40.86±3.13; 43.69±3.09 in moderately differentiated and 47.40±2.31 in poorly differentiated cervical cancer and the difference between poorly differentiated type and other two types has statistical significance. It is 42.39±3.26 in I and 47.08±2.47 in II and the difference between them has statistical significance. PI is 48.80±1.10 in lymph node metastasis and 44.00±3.15 in no lymph node metastasis and the difference has statistical significance. Young patients group is 45.36±2.94 contrasting to 43.63±4.07 in old group and the difference between them has not statistical significance.3 COX-2 proteins expressed at cytolymph and masculine cells were mainly tumor cells, atypical hyperplasia cells and inflammatory reaction cells in biopsy place. Masculine cell is yellow and coloration is different.Gray degree of COX-2 is 104.90±1.72 in normal cervical tissues, 115.05±2.75 in CINI,115.42±3.03 in CIN II, 120.52±1.84 in CINIII and 132.79±2.24 in cervical cancer. In cervical cancer, COX-2 in well differentiated is 130.16+1.65; 133.22±1.36 in moderately differentiated and 134.08±2.11 in poorly differentiated cervical cancer and the difference between poorly differentiated type and other two types has statistical significance. It is 132.24±1.79 in I and 133.63±2.66 in II and the difference between them has no statistical significance. It is 135.70±1.60 in lymph node metastasis and 132.30±1.84 in no lymph node metastasis and the difference has statistical significance. Young patients group is 134.06±2.01 contrasting to 132.06+2.08 in old group and the difference between them has statistical significance.Conclusion1 MVD marked by CD105 related to pathology grade of cervical squamous cell carcinoma and FIGO clinical stage but age of patients. It indicated that angiogenesis related to malignancy degree and scale of cervical squamous cell carcinoma proving tumor angiogenesis. In this study CD105 also related to lymph node metastasis and indicated angiogenesis not only promote scale-up of tumor corresponding to cervical squamous cell carcinoma route of metastasis but also can promote lymph node metastasis.2 Expression of PCNA was related with pathology grade of cervical squamous cell carcinoma, FIGO clinical stage and lymph node metastasis indicating PCNA could reflect proliferate activity of tumor cell. It could be a diagnosis target of cervical squamous cell carcinoma. As CD105 it also has no dependability with age of patients and that indicated extent and malignant degree of cervical squamous cell carcinoma had not statistics difference between different age groups.3 Expression of COX-2 had significant difference in age of patient and lymph node metastasis but in FIGO stage of cervical squamous cell carcinoma. Pathology degree though moderated differentiated had no difference with poorly differentiated. Difference between each CIN and each pathology stage of cervical squamous cell carcinoma were not significant. There is difference in different age groups. May be young patients have more sex behavior then old patients so many young patients have chronic cervicitis and COX-2 has relation with inflammation.4 Expression of CD105, PCNA and COX-2 in normal cervical tissue, cervical intraepithelial neoplasis and cervical squamous cell carcinoma has statistics significance and was direct correlation what hint them play a coordinated role in cervical squamous cell carcinoma. Detection of CD105, PCNA and COX-2 maybe helpful to get the message of biological behavior of cervical squamous cell carcinoma cells to provide the theoretic basis for prevention and therapy in clinic.