本文選題：肝癌 + 腫瘤相關(guān)巨噬細(xì)胞��； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:探討腫瘤相關(guān)巨噬細(xì)胞(tumor-associated macrophages,TAMs)中RIP140的表達(dá)對(duì)巨噬細(xì)胞極化和對(duì)肝癌細(xì)胞侵襲、增殖的影響及其分子機(jī)制。方法:(1)慢病毒介導(dǎo)小鼠腹腔巨噬細(xì)胞(peritoneal macrophages.PMs)RIP140的過(guò)表達(dá),Western blot和Real-time PCR(q RT-PCR)分別檢測(cè)PMs中RIP140蛋白以及核酸表達(dá)水平,流式細(xì)胞儀分析慢病毒轉(zhuǎn)染率;(2)Western blot、細(xì)胞免疫熒光和q RT-PCR檢測(cè)肝癌條件培養(yǎng)基(hepatocellular carcinoma conditioned medium,HCM)刺激PMs24 h后TAMs中RIP140的表達(dá)變化;(3)HCM刺激PMs以及HCM刺激過(guò)表達(dá)RIP140的PMs,q RT-PCR檢測(cè)TAMs極化指標(biāo)以及NF-κB和IL-6的表達(dá),Transwell實(shí)驗(yàn)和細(xì)胞流式凋亡實(shí)驗(yàn)檢測(cè)肝癌細(xì)胞的侵襲和凋亡;(4)肝癌細(xì)胞和PMs以4∶1比例注射于BALB/c裸鼠皮下,建立裸鼠皮下肝癌模型,成瘤癌組織HE染色和免疫組化評(píng)定肝癌組織大體生長(zhǎng)情況和肝癌細(xì)胞增殖能力。結(jié)果:(1)慢病毒介導(dǎo)PMs RIP140的過(guò)表達(dá),病毒轉(zhuǎn)染率較高(約60%),RIP140過(guò)表達(dá)明顯;HCM刺激PMs后,TAMs中RIP140表達(dá)降低;(2)HCM誘導(dǎo)TAMs呈M2型極化,并且與腫瘤生長(zhǎng)密切相關(guān)的NF-κB/IL-6通路處于激活狀態(tài);TAMs可促進(jìn)肝癌細(xì)胞侵襲和增殖,抑制肝癌細(xì)胞凋亡。(3)TAMs過(guò)表達(dá)RIP140可抑制HCM介導(dǎo)的TAMs M2型極化并抑制NF-κB/IL-6通路的激活,減少IL-6的釋放;此外,TAMs過(guò)表達(dá)RIP140可抑制肝癌細(xì)胞的侵襲和增殖[PCNA+細(xì)胞數(shù):對(duì)照組(117.3±13.1)vs過(guò)表達(dá)組(56.9±7.4),P0.05],并促進(jìn)肝癌細(xì)胞的凋亡[凋亡率:對(duì)照組(28.7±3.6%)vs過(guò)表達(dá)組(43.1±2.9%),P0.05]。結(jié)論:過(guò)表達(dá)RIP140的TAMs抑制肝癌細(xì)胞的侵襲和增殖。其機(jī)制與TAMs過(guò)表達(dá)RIP140后抑制TAMs M2型極化有關(guān)。
[Abstract]:Aim: to investigate the effect of RIP140 expression in tumor-associated macrophages (tumor-associated macrophages) on macrophage polarization, invasion and proliferation of hepatoma cells and its molecular mechanism. Methods the overexpression of macrophages.PMs)RIP140 in murine peritoneal macrophages mediated by lentivirus was detected by Western blot and Real-time PCR(q RT-PCR respectively. Flow cytometry analysis of lentivirus transfection rate: Western blot.The cell immunofluorescence and Q RT-PCR were used to detect the expression of RIP140 in TAMs after PMs24 h stimulated by hepatocarcinoma conditioned medium (carcinoma conditioned medium). The RIP140 expression in TAMs was detected by PMs stimulated by PMs and PMs by PMs stimulated by HCM and the TAMs electrode by PMsTQ RT-PCR stimulated by HCM. The expression of NF- 魏 B and IL-6 was detected by Transwell assay and flow cytometry. The invasion and apoptosis of hepatoma cells were detected by transwell assay and flow cytometry. (4) hepatoma cells and PMs were injected subcutaneously into BALB/c nude mice at 4:1. The subcutaneous liver cancer model of nude mice was established, and HE staining and immunohistochemistry were used to evaluate the gross growth and proliferation of hepatoma cells. Results PMs RIP140 overexpression mediated by lentivirus had a high transfection rate (about 60% RIP140 overexpression). The expression of RIP140 in PMs induced by lentivirus-induced TAMs showed M2 polarization. Furthermore, the activation of NF- 魏 B/IL-6 pathway, which is closely related to tumor growth, can promote the invasion and proliferation of hepatocellular carcinoma cells, and inhibit the overexpression of TAMs M2 type polarization mediated by HCM, inhibit the activation of NF- 魏 B/IL-6 pathway and reduce the release of IL-6. In addition, RIP140 overexpression of tams inhibited the invasion and proliferation of HCC cells [PCNA cell number: 56.9 鹵7.4 PCNA cells in the control group: 117.3 鹵13.1)vs overexpression group, P0.05], and promoted apoptosis of HCC cells [apoptosis rate: 43.1 鹵2.9 in the control group, 28.7 鹵3.6%)vs overexpression group, P0.05]. Conclusion: TAMs overexpression of RIP140 inhibits the invasion and proliferation of hepatoma cells. The mechanism is related to the inhibition of M 2 polarization of TAMs after TAMs overexpression of RIP140.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Alexander Schlachterman;Willie W Craft Jr;Eric Hilgenfeldt;Avir Mitra;Roniel Cabrera;;Current and future treatments for hepatocellular carcinoma[J];World Journal of Gastroenterology;2015年28期

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本文編號(hào)：1784682