本文選題：人臍帶間充質(zhì)干細(xì)胞 + 微囊�。� 參考：《上海交通大學(xué)》2014年碩士論文

【摘要】：目的：初步探討人臍帶間充質(zhì)干細(xì)胞來源的微囊（hWJMSC-MVs）對(duì)人腎癌細(xì)胞(RCC）的影響及其可能的作用機(jī)制。 方法：1、選用人腎癌細(xì)胞株786-O作為研究對(duì)象，分三組進(jìn)行干預(yù)：（1）+MVs組，（2）control組，（3）+RNase-MV組（經(jīng)RNA酶處理的MV），24h、48h后檢測(cè)腫瘤細(xì)胞增殖率（CCK-8）及細(xì)胞周期，進(jìn)行劃痕實(shí)驗(yàn)檢測(cè)。運(yùn)用實(shí)時(shí)定量PCR檢測(cè)腎癌細(xì)胞中MMP-2、MMP-9基因的表達(dá)情況。 2、裸鼠皮下注射腎癌細(xì)胞建立腎癌移植瘤模型，每三天測(cè)量一次腫瘤大小，計(jì)算各組腫瘤面積及發(fā)生率，，36d后進(jìn)行犧牲，分別進(jìn)行免疫組化分析（HGF、KI-67及Tunel）。 3、運(yùn)用Western blot技術(shù)檢測(cè)各組腎癌標(biāo)本中MMP-2、MMP-9、cyclinD1的蛋白表達(dá)情況。然后檢測(cè)各組腎癌細(xì)胞或腫瘤標(biāo)本中HGF、c-MET、ERK1/2、AKT蛋白表達(dá)情況，并對(duì)c-MET抑制劑干預(yù)后的腎癌細(xì)胞中p-ERK1/2、p-AKT蛋白表達(dá)情況進(jìn)行檢測(cè)。 結(jié)論：hWJMSC-MVs促進(jìn)了腎癌的生長(zhǎng)及侵襲能力，其作用機(jī)制可能是MVs誘導(dǎo)HGF的增加從而增強(qiáng)了PI3k-AKT及Ras/MAPK（ERK1/2）信號(hào)通路的活化，進(jìn)一步加強(qiáng)了腎癌細(xì)胞增殖及抗凋亡的能力。同時(shí)發(fā)現(xiàn)MV經(jīng)RNA酶處理后喪失了進(jìn)一步促進(jìn)腎癌生長(zhǎng)的能力，證明MVs對(duì)靶細(xì)胞的作用可能是通過傳遞有效的遺傳物質(zhì)（mRNA/miRNA）來實(shí)現(xiàn)的，MVs可能是介導(dǎo)細(xì)胞間相互作用的一種媒介物質(zhì)。本實(shí)驗(yàn)也為HGF/c-MET途徑可能成為腎癌治療的靶點(diǎn)提供了實(shí)驗(yàn)依據(jù)。
[Abstract]:Aim: to investigate the effect of human umbilical cord mesenchymal stem cell derived microcapsules (hWJMSC-MVs) on human renal cell carcinoma (RCC) and its possible mechanism. Methods: 1. Human renal cell carcinoma cell line 786-O was selected as the study object. It was divided into three groups: (1) MVs group, (2) control group, (3) RNase-MV group (RNase-MV) 24 h and 48 h later, the tumor cell proliferation rate (CCK-8) and cell cycle were detected by scratch test. The expression of MMP-2 and MMP-9 gene in renal carcinoma cells was detected by real-time quantitative PCR. 2. The transplanted tumor model was established by subcutaneous injection of renal cancer cells in nude mice, and the tumor size was measured every three days. After 36 days' sacrifice, the tumor area and incidence of each group were analyzed by immunohistochemical analysis (HGFKI-67 and Tunel). Western blot was used to detect the expression of MMP-2, MMP-9 and cyclin D1 in renal cell carcinoma. Then, the expression of HGF- c-METERK1 / 2AK protein in renal cancer cells or tumor samples was detected, and the expression of p-ERK1 / 2 pAKT protein in renal cancer cells after intervention with c-MET inhibitor was detected. Conclusion the growth and invasiveness of RCC were promoted by WJMSC-MVs. The mechanism may be that MVs induces the increase of HGF, which enhances the activation of PI3k-AKT and Rasr-MAPK (ERK1 / 2) signaling pathway, and further strengthens the proliferation and anti-apoptosis ability of RCC cells. At the same time, it was found that MV lost the ability to further promote the growth of renal cell carcinoma after being treated with RNAase, which suggested that MVs' effect on target cells might be mediated by the transmission of effective genetic material (mRNA-miRNA). This study also provides experimental evidence that HGF-c-MET pathway may be a target for the treatment of renal cell carcinoma.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.11

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 米東;張?jiān)孪?陳淑琴;祁s

本文編號(hào)：2100381