The Expression And Significance Of ERCC1 In Renal Clear Cell Carcinoma

Objectives:With the chang of people’s living habits and the improvement modern medical technology, the detection rate of kidney cancer was increasing year by year. now is common in Urinary tract tumors. According to pathology, the kidney cancer was divided into Renal clear cell carcinoma and so on, among them the prevalence of renal clear cell carcinoma is as high as 70 to 80%. recently study found that only early RCCC(T1 ~ 2 stage) can using surgery to servive and the long-term prognosis is good. But for the late RCCC(T3 ~ 4), the efficacy of drugs and surgery are poor. The average life was just 40 months, Therefore develope a new highly specific Index, to improve the diagnosis of renal clear cell cancer, to prolong patients life, is the goal of our research workers have always been.ERCC1 is a link, cut and identify damage genes, ERCC1 was stay in human chromosome 19q13.2 region, the relative molecular weight of 15 k D(1D = 1U). Its m RNA(1.0kb and 1.1kb) are respectively encode 273 and 293 amino acids. Studies have shown that the reason why cells canceration, mainly because of DNA damage can not be repaired in time, eventually leading to chromosomal mutations, nomol cells canceration. Vineis [6] found that the action of ERCC1 gene is identifying and repairing damaged DNA in the development of cancer. so ERCC1 as a key repair damage gene received extensive attention, The expression of ERCC1 in RCCC have never been found.In this study, using IHC and RT-PCR research the expression of ERCC1 in human RCCC and normal kidney tissue, Explore its role in cancer arising from the occurrence and development.Methods:Select 50 cases of renal clear cell carcinoma patients which live in the Second Hospital of Hebei Medical University urology clinic from November 2013 to May 2014 as research objects. Among them men accounted for 28 cases, women accounted for 22 cases, aged 35 to 78 years, mean(54.40 ± 9.56) years. All of the patients were not use radiotherapy and chemotherapy before surgery. Select 50 renal clear cell carcinoma tissue as the patient group. 20 cases of adjacent tissues(paracancer> 3cm of normal kidney tissue) as a common group.one group organization fixed by 4% formaldehyde, then using paraffin embedded the tissue, then cut them into 4μm thin slices, put another group into liquid nitrogen to storage. According to the 1997 release of the WHO Fuhrman three classification criteria for pathological grade.Immunohistochemical technique: Using ICH detected the ERCC1 which in paraffin-embedded tissues from protein levels.There were 50 RCCC tissue samples and 20 normal renal tissues samples. The proportion of cells expressing the number of applications in the light microscope and expression of positive intensity score the results of its comprehensive determination. Compare ERCC1 the differences of the positive expression rate between RCCC and normal kidney tissues. Analysis the relasionship of ERCC1 and histological grade, age, gender.RT-PCR technique: the strict organization of the RNA extraction kit according to c DNA using the method described, the reaction system was added RT tube, disposed within the GAPDH gene fragment as reference gene ERCC1 gene expression, in for the next 94 ℃ thermostatic denaturing conditions 3min PCR amplification primers amplified specimens out of the product line by agarose gel electrophoresis, a 1% agarose gel containing GV nucleic acid dye and DNA Marker(DL2000) for fragment identifies its guidelines by UV transilluminator after electrophoresis observation, computer applications in the input image analysis software Quantity One gel electrophoresis for the purpose of routine analysis with rows within the selection of the appropriate internal control reference electrophoretic bands, results in both the ratio of the integrated absorbance of representation. Methods: Absorbance ratio between the two groups using the t test was used for statistical analysis.Using SPSS17.0 statistical software to analysis all of the statistical data. If P <0.05 the difference had statistically significant.Results:1 Immunohistochemical staining showed: The expression of ERCC1 were both in RCCC and adjacent normal renal tissues, The positive expression rate in RCCC was 60% higher than which in normal tissues 25%; the expression of ERCC1 in well-differentiated and middle- differentiated group more than poorly differentiated group(χ2 = 8.680, P <0.05), The expression of ERCC1 were not related with age and gender(P > 0.05).2 RT-PCR results showed that: The expression of ERCC1 in renal clear cell carcinoma was 0.280 ± 0.030, whereas the expression of ERCC1 in the adjacent normal tissue was 0.104 ± 0.011.Conclusions:1 The expression of ERCC1 in RCCC is higher than the normal tissues, which adjacent to the cancer.2 The expression of ERCC1 had affect in the pathological differentiation of RCCC.3 The age and gender had no effect on the expression of ERCC1.4 The highly expressed ERCC1 protein in RCCC plays an important role in the development of the cancer, future may become an important indicator to judge the progress of the tumor.