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.
【摘要】目的通過RNA干擾(RNAi)沉默HER2基因在涎腺黏液表皮樣癌Mc3細胞中的表達方法將目的基因靶序列小干擾RNA(siRNA)轉染Mc3細胞,并設置對照組,采用RTPCR、免疫組化檢測RNA干擾后HER2基因在Mc3細胞中的表達情況。結果RTPCR結果顯示RNA干擾后,HER2基因mRNA在涎腺黏液表皮樣癌細胞中的表達與對照組比較明顯降低;免疫組化實驗結果顯示RNA干擾后HER2基因蛋白在涎腺黏液表皮樣癌細胞中的表達降低,與mRNA表達情況相一致。結論RNA干擾成功抑制了涎腺黏液表皮樣癌細胞中HER2基因的表達,為口腔涎腺黏液表皮樣癌針對癌基因HER2為靶基因的基因治療提供研究基礎。
目的：研究青蒿琥酯誘導人胃癌SGC-7901細胞凋亡作用及其可能機制.方法：應用流式細胞術（FCM）、透射電鏡（TEM）等方法檢測不同濃度下青蒿琥酯對人胃癌SGC-7901細胞生長的影響，并應用Western Blot法檢測凋亡相關基因Bcl-2、Bax的表達水平。結果：經青蒿琥酯處理后，人胃癌SGC-7901細胞凋亡率升高，并具有時間濃度依賴性（Plt;0.05），癌細胞被阻滯于G0/G1期；電鏡觀察腫瘤組織中散在凋亡細胞及凋亡小體；Western Blot法檢測到凋亡相關基因Bcl-2表達減弱，Bax表達增強（Plt;0.05）。結論：青蒿琥酯能有效抑制人胃癌SGC7901細胞增殖并誘導其凋亡，其機制可能與下調凋亡相關基因Bcl-2、上調Bax表達有關。
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)