目的:探討KAI1基因敲低表達(dá)后對(duì)肺腺癌細(xì)胞生物學(xué)行為和功能的影響以及和腫瘤信號(hào)相關(guān)通路分子之間的關(guān)系。方法:構(gòu)建目的基因KAI1基因敲低表達(dá)組和陰性對(duì)照組干擾片段siRNA,利用瞬時(shí)轉(zhuǎn)染技術(shù)通過轉(zhuǎn)染試劑Lpofectamin2000作用于兩種肺腺癌細(xì)胞株A549和H1299,利用RT-PCR技術(shù)和Western blot技術(shù)在基因和蛋白水平分別檢測目的基因在KAI1空白組、陰性對(duì)照組、敲低表達(dá)組的表達(dá);MTT試劑盒法以及Transwell小室侵襲和遷移技術(shù)檢測KAI1基因敲低表達(dá)后肺腺癌細(xì)胞功能變化情況;利用Western blot檢測KAI1基因敲低表達(dá)后Axin1、GSK3β、vimentin和Slug的蛋白表達(dá)水平在各組中的變化情況;將慢病毒介導(dǎo)的KAI1過表達(dá)基因轉(zhuǎn)染到人肺腺癌細(xì)胞A549后注射到裸鼠體內(nèi)觀測其對(duì)裸鼠移植瘤生長的影響,最后收集每組數(shù)據(jù)計(jì)算移植瘤的平均體積,連續(xù)觀察5周后脫頸處死裸鼠并剝離瘤組織,并稱取瘤組織的重量、測量瘤體體積。結(jié)果:兩株細(xì)胞KAI1敲低表達(dá)組中的KAI1基因mRNA表達(dá)水平均比空白組和陰性對(duì)照組低,數(shù)據(jù)差異具有統(tǒng)計(jì)學(xué)意義（p<0.05）... 

【文章來源】：蚌埠醫(yī)學(xué)院安徽省

【文章頁數(shù)】：62 頁

【學(xué)位級(jí)別】：碩士

【部分圖文】：

熒光定量PCR檢測兩株細(xì)胞KAI1mRNA的表達(dá)Figure1:FluorescencequantitativePCRdetectionofKAI1mRNAexpressionintwocells

22圖1：熒光定量PCR檢測兩株細(xì)胞KAI1mRNA的表達(dá)Figure1:FluorescencequantitativePCRdetectionofKAI1mRNAexpressionintwocells圖2：兩株細(xì)胞干擾組中KAI1mRNA的相對(duì)表達(dá)量明顯低于空白組和NC組（p＜0.05）Figure2:TherelativeexpressionofKAI1mRNAintwocellinterferencegroupswassignificantlylowerthanthatinblankgroupandNCgroup(p<0.05)

KAI1 KAI1 Tubulin Tubulin 圖 3：兩株細(xì)胞各實(shí)驗(yàn)組蛋白表達(dá) Figure 3: Protein expression in each experimental group of two cells

【參考文獻(xiàn)】：
期刊論文
[1]Expression and clinical significance of p53,JunB and KAI1/CD82 in human hepatocellular carcinoma[J]. Cheng Guo,Qing-Guang Liu,Lei Zhang,Tao Song and Xue YangDepartment of Hepatobiliary Surgery,First Affiliated Hospital,School of Medicine,Xi’an Jiaotong University,Xi’an 710061, China.  Hepatobiliary & Pancreatic Diseases International. 2009(04)
[2]KAI1 gene expression in colonic carcinoma and its clinical significances[J]. De-Hua Wu Li Liu Long-Hua Chen Yan-Qing Ding Department of Radiation Oncology,Nanfang Hospital,First Military Medical University,Guangzhou 510515,Guangdong Province,ChinaDepartment of Pathology,First Military Medical University,Guangzhou 510515,Guangdong Province,China.  World Journal of Gastroenterology. 2004(15)



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