本文關(guān)鍵詞： 核心蛋白聚糖 肝癌 P21~(WAF1/CIP1) TGF-β_1 caspase-3 caspase-8　出處：《吉林大學(xué)》2008年博士論文　論文類型：學(xué)位論文

【摘要】： 核心蛋白聚糖(decorin,DCN)屬于分泌型、小分子、富含亮氨酸多糖基因擴(kuò)展家族成員之一,主要存在于結(jié)締組織中并且是一種與膠原纖維相關(guān)的蛋白多糖,研究發(fā)現(xiàn)它能抑制細(xì)胞增殖和抗纖維化,具有抗腫瘤的作用。目前國(guó)內(nèi)外還沒(méi)有decorin基因轉(zhuǎn)染到肝癌細(xì)胞HepG2中以及研究decorin抗肝癌機(jī)制的報(bào)道,本實(shí)驗(yàn)采用體外基因重組技術(shù)構(gòu)建了真核表達(dá)載體pcDNA3.1-decorin,并且轉(zhuǎn)染到肝癌細(xì)胞HepG2中,探討其對(duì)肝癌細(xì)胞的作用機(jī)理。本研究成功構(gòu)建了真核表達(dá)載體pcDNA3.1-decorin,并轉(zhuǎn)染到肝癌細(xì)胞HepG2中;結(jié)果發(fā)現(xiàn)decorin能夠抑制肝癌細(xì)胞HepG2生長(zhǎng),FCM方法檢測(cè)細(xì)胞周期發(fā)現(xiàn)pcDNA3.1-dec-HepG2組G0/G1期細(xì)胞比pcDNA3.1-HepG2組G0/G1期細(xì)胞明顯升高、S期細(xì)胞顯著降低,細(xì)胞生長(zhǎng)停滯在G0/G1期; Caspase-3、Caspase-8活性檢測(cè)發(fā)現(xiàn)pcDNA3.1-dec-HepG2組細(xì)胞比pcDNA3.1-HepG2組細(xì)胞顯著升高,RT-PCR和western blot方法顯示pcDNA3.1-dec-HepG2組細(xì)胞周期依賴激酶抑制劑P21 mRNA和蛋白質(zhì)比pcDNA3.1-HepG2組細(xì)胞顯著升高, RT-PCR方法顯示pcDNA3.1-dec-HepG2組細(xì)胞轉(zhuǎn)化生長(zhǎng)因子TGF-β1比pcDNA3.1-HepG2組細(xì)胞顯著降低。上述結(jié)果為進(jìn)一步研究decorin抗腫瘤的作用機(jī)制奠定了基礎(chǔ),同時(shí)也為decorin的臨床應(yīng)用提供了理論依據(jù)。
[Abstract]:Core protein decorin (DCNs) belongs to secretory type, small molecule, rich in leucine polysaccharide gene expansion family members. It is mainly found in connective tissue and is a proteoglycan associated with collagen fiber. It has been found that it can inhibit cell proliferation and resist fibrosis. At present, there are no reports of decorin gene transfection into HCC HepG2 and study of the mechanism of decorin anti-HCC. In this study, eukaryotic expression vector pcDNA3.1-decorin was constructed by in vitro gene recombination technique and transfected into hepatoma cell HepG2. In this study, eukaryotic expression vector pcDNA3.1-decorin was constructed and transfected into hepatoma cell HepG2. The results showed that decorin could inhibit the growth of HepG2 cells. FCM assay showed that G0 / G1 phase cells in pcDNA3.1-dec-HepG2 group were significantly higher than those in pcDNA3.1-HepG2 group. S phase cells decreased significantly, cell growth stopped at G 0 / G 1 phase. The activity of Caspase-3 and Caspase-8 in pcDNA3.1-dec-HepG2 group was significantly higher than that in pcDNA3.1-HepG2 group. Expression of cell cycle dependent kinase inhibitor P21 in pcDNA3.1-dec-HepG2 group by RT-PCR and western blot. MRNA and protein were significantly higher than those in pcDNA3.1-HepG2 group. RT-PCR method showed that TGF- 尾 1 in pcDNA3.1-dec-HepG2 group was significantly lower than that in pcDNA3.1-HepG2 group. It lays a foundation for further study on the mechanism of anti-tumor effect of decorin. It also provides a theoretical basis for the clinical application of decorin.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2008
【分類號(hào)】：R346

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 葛秀君;劉志輝;高瑞萍;;子宮內(nèi)膜癌p21WAF1/CIP1蛋白的表達(dá)及其意義[J];癌變.畸變.突變;2007年04期

2 趙春霞,趙蔭濤,汪培華,肖嘯,汪道文;重組腺相關(guān)病毒介導(dǎo)核心蛋白聚糖基因轉(zhuǎn)染對(duì)SiHa細(xì)胞周期和凋亡的影響[J];癌癥;2005年01期

3 操海萍;舒振波;張桂珍;;人核心蛋白聚糖真核表達(dá)載體的構(gòu)建及鑒定[J];吉林大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2006年04期

4 張曉暉,姚天明,黃高f,王文亮;細(xì)胞凋亡的最新研究進(jìn)展[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2002年S1期

5 趙素蓮,王桂琴,田樹華;Decorin對(duì)肝癌細(xì)胞株凋亡作用的研究[J];腫瘤防治雜志;2002年03期

6 王慧君,張志剛,陳廣平,林文生,陳琦,郭慕依;穩(wěn)定表達(dá)飾膠蛋白聚糖基因的大鼠腎小球系膜細(xì)胞株的建立[J];復(fù)旦學(xué)報(bào)(醫(yī)學(xué)科學(xué)版);2001年02期

7 戰(zhàn)雪梅,王國(guó)新,孫崇偉,王家銀,李玲;食管癌組織中p16、p21、p53cyclinD1表達(dá)及其意義[J];實(shí)用癌癥雜志;2001年01期

8 王艷紅;王桂琴;郝小夏;;核心蛋白聚糖原核表達(dá)體系的構(gòu)建[J];中國(guó)藥物與臨床;2006年03期

9 邢國(guó)宏,張進(jìn)川;基因芯片檢測(cè)Ⅰ期低分化肺腺癌基因表達(dá)譜[J];中華腫瘤雜志;2004年01期

10 郭新珍,李懷斌,楊玉秀,劉正國(guó);TGF-β1、TβRⅡ及cyclinD1在大腸癌的表達(dá)及其意義[J];腫瘤防治研究;2005年01期

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