發(fā)布時(shí)間：2018-04-15 14:05

  本文選題：CBX8 + 肝細(xì)胞肝癌��； 參考：《青島大學(xué)》2017年碩士論文

【摘要】：目的通過檢測CBX8 m RNA和蛋白質(zhì)在肝細(xì)胞肝癌組織中的表達(dá)及對肝癌細(xì)胞系SMMC-7721增殖、侵襲能力和細(xì)胞周期的影響,初步探討CBX8在肝細(xì)胞肝癌發(fā)生發(fā)展中的作用及其意義。方法懫用實(shí)時(shí)熒光定量PCR和western blot分別檢測15對新鮮肝細(xì)胞肝癌手術(shù)標(biāo)本及癌旁組織中CBX8 m RNA和蛋白質(zhì)的表達(dá);懫用組織芯片技術(shù)及免疫組織化學(xué)方法檢測CBX8蛋白質(zhì)在120例肝細(xì)胞肝癌手術(shù)標(biāo)本及癌旁組織中的表達(dá)情況;分析CBX8的表達(dá)與肝癌發(fā)生及臨床病理特征之間的關(guān)系。分別在肝癌細(xì)胞系SMMC-7721中瞬時(shí)轉(zhuǎn)染小干擾RNA和過表達(dá)質(zhì)粒,通過MTT、平板克隆形成實(shí)驗(yàn)及transwell小室檢測CBX8的敲降和過表達(dá)對肝癌細(xì)胞系SMMC-7721細(xì)胞增殖及侵襲能力的影響,通過細(xì)胞同步化及流式細(xì)胞儀檢測CBX8對肝癌細(xì)胞系SMMC-7721細(xì)胞周期的影響。結(jié)果實(shí)時(shí)熒光定量PCR結(jié)果顯示CBX8 m RNA在13例(13/15)新鮮肝細(xì)胞肝癌手術(shù)標(biāo)本中表達(dá)量增高,15例肝細(xì)胞肝癌與癌旁組織中的差異具有統(tǒng)計(jì)學(xué)意義(t=2.742,P0.05)。Western blot結(jié)果表明CBX8蛋白質(zhì)在11例(11/15)新鮮肝細(xì)胞肝癌組織中表達(dá)量增高,15例肝細(xì)胞肝癌與癌旁組織中的差異具有統(tǒng)計(jì)學(xué)意義(t=2.686,P0.05)。組織芯片及免疫組化檢測結(jié)果顯示CBX8在100例(83.33%)肝細(xì)胞肝癌手術(shù)標(biāo)本中表達(dá),在46例(38.33%)癌旁組織中表達(dá),CBX8在120例肝細(xì)胞肝癌及癌旁組織的表達(dá)差異具有統(tǒng)計(jì)學(xué)意義(Z=-8.072,P0.05)。CBX8在肝細(xì)胞肝癌中的表達(dá)水平與患者乙肝表面抗原(HBs Ag)、腫物直徑、被膜侵犯以及TNM分期有關(guān)(?2=10.900、4.043、46.570、8.374,P0.05)。敲降CBX8后,肝癌細(xì)胞系SMMC-7721細(xì)胞的增殖、克隆形成率和遷移侵襲能力均下降,且與對照組相比差異均具有統(tǒng)計(jì)學(xué)意義(F=9.395、24.300、71.180、8.745、63.700、9.044,P0.05);過表達(dá)CBX8則結(jié)果相反,并且與對照組相比差異均具有統(tǒng)計(jì)學(xué)意義(F=7.177、36.100、24.680、9.045、7.037、6.525,P0.05)。通過檢測細(xì)胞周期,我們發(fā)現(xiàn)敲降CBX8后,實(shí)驗(yàn)組G1期細(xì)胞所占比例明顯增加,S期細(xì)胞所占比例減少,差異有統(tǒng)計(jì)學(xué)意義(t=3.170、3.269,P0.05);過表達(dá)CBX8后,實(shí)驗(yàn)組G1期細(xì)胞所占比例明顯減少,S期細(xì)胞所占比例增加,差異有統(tǒng)計(jì)學(xué)意義(t=2.884、4.443,P0.05)。結(jié)論CBX8在肝細(xì)胞肝癌中呈顯著高表達(dá);CBX8在肝細(xì)胞肝癌的發(fā)生發(fā)展中發(fā)揮重要作用,能夠促進(jìn)肝癌細(xì)胞系SMMC-7721的增殖、遷移、侵襲等生物學(xué)行為,并可能通過調(diào)控細(xì)胞周期G1/S轉(zhuǎn)換進(jìn)而影響SMMC-7721細(xì)胞周期進(jìn)程;CBX8在肝細(xì)胞肝癌發(fā)生發(fā)展中發(fā)揮癌基因作用。
[Abstract]:Objective to investigate the effect of CBX8 m RNA and protein on the proliferation, invasion and cell cycle of hepatocellular carcinoma (HCC) cell line SMMC-7721 by detecting the expression of CBX8 m RNA and protein in HCC tissues.Methods Real-time quantitative PCR and western blot were used to detect the expression of CBX8 m RNA and protein in 15 fresh hepatocellular carcinoma specimens and adjacent tissues.Tissue microarray technique and immunohistochemical method were used to detect the expression of CBX8 protein in 120 HCC specimens and adjacent tissues, and the relationship between the expression of CBX8 and the occurrence and clinicopathological features of HCC was analyzed.Transient transfection of small interfering RNA and overexpression plasmids were carried out in hepatocellular carcinoma (SMMC-7721) cell lines. The effects of CBX8 knockdown and overexpression on the proliferation and invasiveness of SMMC-7721 cells were detected by MTT, plate clone formation assay and transwell chamber.Cell synchronization and flow cytometry were used to detect the effect of CBX8 on the cell cycle of hepatocellular carcinoma cell line SMMC-7721.Results the results of real-time fluorescence quantitative PCR showed that the expression of CBX8 m RNA was increased in 13 cases of fresh hepatocellular carcinoma and 15 cases of hepatocellular carcinoma. The difference between 15 cases of hepatocellular carcinoma and adjacent tissues was statistically significant. Western blot showed that CBX8 protein was expressed in 13 cases of fresh hepatocellular carcinoma.In 11 cases of fresh hepatocellular carcinoma (11 / 15), there was a significant difference between 15 cases of hepatocellular carcinoma (HCC) and adjacent tissues (P 0.05).The results of tissue microarray and immunohistochemistry showed that CBX8 was expressed in 100 specimens of hepatocellular carcinoma (HCC).There were significant differences in the expression of CBX8 in 120 cases of hepatocellular carcinoma (HCC) and its adjacent tissues in 46 cases (38.33). The expression level of CBX8 in hepatocellular carcinoma (HCC) was significantly higher than that in patients with Hepatocellular carcinoma (HCC).The membrane invasion and TNM staging were related to 46.570 ~ 8.374g / L P0.05A, P = 10.900 ~ 4.043n ~ 46.570g / L, P = 0.05a, P = 0.05a, respectively.After knockdown of CBX8, the proliferation, clone formation rate and migration and invasion ability of SMMC-7721 cell line decreased, and the difference was statistically significant compared with the control group. The difference was statistically significant compared with the control group.Compared with the control group, the difference was statistically significant (P 0.05).By measuring cell cycle, we found that the percentage of G 1 phase cells in the experimental group decreased significantly after knocking down CBX8, and the difference was statistically significant (P 0.05) after overexpression of CBX8.The percentage of G 1 phase cells in the experimental group was significantly decreased and the percentage of S phase cells was increased. The difference was statistically significant (P 0.05).Conclusion CBX8 plays an important role in the carcinogenesis and development of hepatocellular carcinoma (HCC), and can promote the proliferation, migration, invasion and other biological behaviors of HCC cell line SMMC-7721.CBX8 may play an oncogene role in the carcinogenesis and development of hepatocellular carcinoma (HCC) by regulating the G1 / S transition of cell cycle and then affecting the progression of SMMC-7721 cell cycle.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.7

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