Objective To compare gene express difference ofkeloid and normal skin tissues by using the suppression subtractive hybridization (SSH) so asto find the differential express gene in keloid. Methods mRNA extracted fromkeloid and normal skin tissues was used as the template to synthesis cDNA of keoid and normal skin. The cDNA of keloid served as a tester, the cDNA of normal skin as a driver. cDNA was digested with RsaⅠ. Adaptor-ligated tester cDNA was prepared. Then first hybridization, second hybridization and PCR amplificationwere done. Differentially expressed cDNA was selectively amplified during thesereactions. After SSH, the PCR mixture was ligated with T-vector. The positive clones were selected and the insert gene fragments were analyzed. Southern hybridization identified the keloid differential express genes. The positive clones ofSouthern hybridization were selected, and these sequences were analyzed. The results were compared with that of GeneBank. Results Thirteen differential genes were found in keloid, of which 11 gene clones have been known their function, and 2 clones have not known their function. 〖WTHZ〗Conclusion Keloid differentially expressed gene was screened successfully by SSH.
Purpose To investigate nucleoside diphosphate kinase (NDPK ) expression of tumor metastasis suppressor gene nm23 in heterotransplanted model of retinoblastoma(RB) in nude mice，and analyse the correlation between the expression of nm23 gene and the formation and progression of heterotran splanted RB． Methods SP immunohistochemical method was used to detect the expression of nm23 gene product NDPK in 20 tumors of heter otransplanted RB model and normal retinal tissue． Results The negative staining of nm23/ NDPK was found in normal retinal tissue , whereas 100% expression rate in RB tumors with positive number of 48.73plusmn;2.37． No statistical significance of the expression of nm23/ NDPK was observed between the intraocular growth phase （I～Ⅲ grade） and invasive phase （ Ⅳ～Ⅴ grade）in heterotransplantedRB tumors． Conclusion The function of nm23 gene as a tumor metastasis suppressor in heterotra nsplanted RB tumors was less prominent ,but it may play a role in carcinogen esis and progrssion of RB and may predict poor prognosis． (Chin J Ocul Fundus Dis, 2001.17:47-49)
【摘要】目的通过RNA干扰(RNAi)沉默HER2基因在涎腺黏液表皮样癌Mc3细胞中的表达方法将目的基因靶序列小干扰RNA(siRNA)转染Mc3细胞,并设置对照组,采用RTPCR、免疫组化检测RNA干扰后HER2基因在Mc3细胞中的表达情况。结果RTPCR结果显示RNA干扰后,HER2基因mRNA在涎腺黏液表皮样癌细胞中的表达与对照组比较明显降低;免疫组化实验结果显示RNA干扰后HER2基因蛋白在涎腺黏液表皮样癌细胞中的表达降低,与mRNA表达情况相一致。结论RNA干扰成功抑制了涎腺黏液表皮样癌细胞中HER2基因的表达,为口腔涎腺黏液表皮样癌针对癌基因HER2为靶基因的基因治疗提供研究基础。
目的：研究青蒿琥酯诱导人胃癌SGC-7901细胞凋亡作用及其可能机制.方法：应用流式细胞术（FCM）、透射电镜（TEM）等方法检测不同浓度下青蒿琥酯对人胃癌SGC-7901细胞生长的影响，并应用Western Blot法检测凋亡相关基因Bcl-2、Bax的表达水平。结果：经青蒿琥酯处理后，人胃癌SGC-7901细胞凋亡率升高，并具有时间浓度依赖性（Plt;0.05），癌细胞被阻滞于G0/G1期；电镜观察肿瘤组织中散在凋亡细胞及凋亡小体；Western Blot法检测到凋亡相关基因Bcl-2表达减弱，Bax表达增强（Plt;0.05）。结论：青蒿琥酯能有效抑制人胃癌SGC7901细胞增殖并诱导其凋亡，其机制可能与下调凋亡相关基因Bcl-2、上调Bax表达有关。