人乳头瘤病毒16型E7蛋白表达对RMA细胞在体内外生长的影响(英文)

Objective:The aim of this study was to study the effect of human papilloma virus(HPV) type 16 E7 protein expression on growth of RMA cells in vitro and in vivo.Methods:The recombination vector pcDNA3.1-E7 carrying wild type HPV 16 E7 was identified by sequencing.The recombination vector pcDNA3.1-E7 was transfected into mouse lymphadenoma cell line RMA by liposome,and the monoclonal cells transfected stably were obtained by antibiotics G418 sieving and limiting dilution assay.RT-PCR method was used to detect the expression of HPV 16 E7 mRNA in RMA-E7 cells.The growth of RMA cells and RMA-E7 cells cultured in vitro was tested by Cell Count Kit-8.RMA-E7 cells and RMA cells were subcutaneously inoculated in syngeneic mice respectively,the tumor size was measured by sliding caliper twice a week,and the E7 protein expression in tumor tissue of mice was detected by Western blot after tumor formation.The kinetics of cytolytic activity of E7 specific T cells in tumor-bearing mice was measured by LDH kit.Results:Sequencing of recombination vector showed the target gene which was inserted into the recombinant was correct,and RMA-E7 cells expressing E7 protein stably were obtained by limited dilution assay.There were no obvious differences in morphous and growth velocity between RMA cells and RMA-E7 cells in vitro.RMA-E7 cells grew in syngeneic mice were significantly slower than RMA cells.The E7 protein was expressed stronger in RMA-E7 cells in vivo than in vitro.The cytolytic ability of E7-specific CTL was activated at the early stage,reached the maximum at the middle stage,and lost at the end stage.RMA-E7 cells isolated from the tumor-bearing mice were more resistant to E7-specific CTL killing than RMA-E7 cells cultured in vitro.Conclusion:The E7 protein expression has no obvious influence on growth of RMA-E7 cells in vitro,and can suppress growth of RMA-E7 cells in vivo.The activity curve of E7 specific CTL approximately presents "bell" shape.The RMA-E7 cells grew in vivo had a high expression levels of E7 protein,and more resistant to E7-specific CTL killing than those cultured in vitro.The E7 protein expression in vivo not only initiates immune activation,but also induces immune tolerance.

Expression of Human Papillomavirus in Mammary Carcinoma and its Possible Mechanism in Carcinogenesis

To explore the role of Human papillomavirus (HPV) in mammary carcinogenesis, the expression of the HPV-16, iNOS , P53 and hTERT proteins in breast carcinomas and their relationships were investigated. 52 samples of breast cancer and 16 samples of benign breast tumors were assayed using the immunohistochemical SP method for detection of protein expression levels. The expression of HPV-16, iNOS, P53 and hTERT proteins in a mammary carcinoma was 44.2%, 57.7%, 63.5% and 59.6% respectively, which was significantly greater than the corresponding levels in the benign group. The expression of iNOS, P53 and hTERT was correlated with the presence of an HPV-16 infection in a mammary carcinom(aP<0.05). The connection between these events might also involve the iNOS, mutated type P53 and the hTERT protein.

Expression of E2F-1, Rb and ER in Peripheral Papil-loma and Ductal Carcinoma in Situ of the Breast and its Significance

OBJECTIVE To investigate the correlation of E2F-1, Rb and ER expression with peripheral papilloma (Peri-PM) and ductal carcinoma in situ of the breast (DCIS), and further explore some molecular mechanisms of the canceration of Peri-PM. METHODS Imunohistochemistry was used to examine the expression of E2F-1, Rb and ER in 60 Peri-PM, 60 Peri-PM with atypical ductal hyperplasia (Peri-PM with ADH) and 60 DCIS. Normal breast tissues were selected as a control group. RESULTS Based on immunohistochemical staining, the positive rate of E2F-1 expression in Peri-PM, Peri-PM with ADH and DCIS was 21.7%, 46.7% and 78.3% respectively. The positive rate of Rb expression was 83.3 %, 53.9% and 21.7% and the ER expression was 86.7%,61.7% and 55.0%. Significant differences were found among the 3 groups (Peri-PM, Peri-PM with ADH and DCIS) (P<0.05). Significant differences existed between any 2 groups (P<0.05) except for the rate of ER positive expression comparing Peri-PM with ADH verus DCIS (P>0.05). The expression of E2F-1 was nega- tively correlated with ER and Rb, and at the same time the expression of ER was positively correlated with Rb. Following the degree of breast epithelial hyperplasia involved and its development into carcinoma, the positive rate of E2F-1 expression displayed an elevating tendency, but that of Rb and ER expression showed a tendency to decline. CONCLUSION The interaction of the 3 indexes studied may play an im- portant role in the conversion of precancerous lesions to early in situ breast carcinoma, and the evaluation of these indexes might provide a valuable basis for screening high-risk cases of Peri-PM.

Study of Benign and Malignant Intraductal Lesions of the Breast by Fiberoptic Ductoscopy

OBJECTIVE To observe and subtype the appearance of intraductal papilloma (lesions) and of infiltrating ductal carcinoma or early infiltrating ductal carcinoma using a fiberoptic ductoscope (FDS) examination, and to discuss the differentiation and diagnosis of benign and malignant tumors,by FDS.METHODS The characteristics of FDS images and diagnostic data for 229 patients with intraductal papilloma (lesions) and 50 patients with ductal carcinoma, who were confirmed by surgical pathology from October 1998 to December 2003, were analyzed retrospectively.RESULTS The appearance of the lesions observed by FDS were grouped into 4types: a monothelia (type Ⅰ), polythelia (type Ⅱ), superficies (type Ⅲ) and a mixture (type Ⅳ). Intraductal papillomas (lesions) were more commonly seen in type Ⅰ and Ⅱ, and intraductal carcinomas, or early infiltrating ductal carcinomas were more commonly seen in type Ⅲ and Ⅳ;there was a statistically significant difference in the distribution of the ductoscopic types, except in type Ⅱ, between the two types of lesions, P<0.001. The focal detection rate by FDS for intraductal papilloma and papillomatosis was 99.6% (228/229) and for breast cancer was 96.0% (48/50). The diagnostic accuracy was 97.8% (224/229) and 82.0% (41/50),respectively.CONCLUSION FDS can be a guide for the treament of benign and malignant intraductal tumors, with early discovery and accurate diagnosis.