本文選題：卵巢癌 + 羅勒多糖�。� 參考：《山東大學》2014年碩士論文

【摘要】：背景與目的卵巢癌是女性生殖系統(tǒng)常見惡性腫瘤之一,其死亡率居于婦科惡性腫瘤的首位；腫瘤侵襲轉移是致死的主要原因。目前手術、放療及化療仍是卵巢癌的主要治療手段,但其療效仍相對有限。部分傳統(tǒng)中醫(yī)藥在惡性腫瘤治療中展現出較好的療效和獨特的優(yōu)勢；本論文主要研究傳統(tǒng)活血化瘀方成分之一的提取物羅勒多糖對卵巢癌SKOV3細胞侵襲能力的影響,并從羅勒多糖對血管內皮生長因子受體表達的調控作用這一角度探討其可能的參與機制。 方法Transwell侵襲試驗研究、檢測不同濃度羅勒多糖對SKOV3細胞侵襲能力的影響；酶聯免疫吸附試驗(ELISA)檢測羅勒多糖對SKOV3細胞分泌血管內皮生長因子(VEGF)的影響,實時定量PCR (qRT-PCR)、流式細胞術(FCM)檢測羅勒多糖對VEGF受體(VEGFRs)在SKOV3細胞表達的調控作用；設計針對VEGFR2/KDR的shRNA及陰性對照shRNA,并構建攜帶不同shRNA的pLL3.7慢病毒表達載體,采用293T細胞包裝攜帶shRNA的慢病毒；慢病毒感染SKOV3細胞后,流式細胞術分選其中的GFP陽性細胞,qRT-PCR、流式細胞術驗證shRNA對VEGFR2表達的敲減作用；Transwell試驗檢測穩(wěn)定敲減VEGFR2在SKOV3細胞表達對羅勒多糖侵襲抑制能力的影響。結果Transwell侵襲試驗表明羅勒多糖可顯著抑制SKOV3細胞的體外侵襲能力,并具有一定的濃度依賴性；羅勒多糖并未影響SKOV3細胞VEGF分泌水平的表達,對VEGFR1、VEGFR3在SKOV3的表達也無顯著影響,但可明顯上調VEGFR2在SKOV3細胞mRNA水平和膜表面蛋白水平的表達；成功感染穩(wěn)定表達針對VEGFR2/KDR shRNA的慢病毒后,qRT-PCR及流式細胞術檢測結果表明VEGFR2在SKOV3細胞的表達顯著下調；進一步研究發(fā)現羅勒多糖對敲減VEGFR2表達的SKOV3細胞侵襲抑制能力顯著降低。 結論羅勒多糖這一傳統(tǒng)中藥提取物可顯著抑制卵巢癌細胞SKOV3細胞的侵襲能力,其抑制作用部分是通過上調VEGFR2在SKOV3細胞的表達介導的。
[Abstract]:Background & objective Ovarian cancer is one of the most common malignant tumors in female reproductive system. Surgery, radiotherapy and chemotherapy are still the main treatment for ovarian cancer, but the efficacy is still relatively limited. Part of traditional Chinese medicine has shown good curative effect and unique advantage in the treatment of malignant tumor. This paper mainly studied the effect of basil polysaccharide, one of the components of traditional Huoxue Huayu recipe, on the invasion ability of ovarian cancer SKOV3 cells. From the perspective of the regulation of basil polysaccharide on the expression of vascular endothelial growth factor receptor, the possible mechanism of its involvement was discussed. Methods Transwell invasion assay was used to detect the effect of basil polysaccharides at different concentrations on the invasiveness of SKOV3 cells, and the effects of basil polysaccharides on the secretion of vascular endothelial growth factor (VEGF) in SKOV3 cells were detected by Elisa assay. The regulation of basil polysaccharides on the expression of VEGF receptor VEGFRsin SKOV3 cells was detected by real-time quantitative PCR qRT-PCRG and flow cytometry, shRNA targeting VEGFR2/KDR and negative control shRNAs were designed, and pLL3.7 lentivirus expression vectors carrying different shRNA were constructed. The lentivirus carrying shRNA was packaged with 293T cells. After lentivirus was infected with SKOV3 cells, Flow cytometry (FCM) was used to detect the inhibitory effect of shRNA on VEGFR2 expression in SKOV3 cells. Flow cytometry was used to detect the effect of stable knockout VEGFR2 expression on the invasion inhibition of basil polysaccharides in SKOV3 cells. Results Transwell invasion test showed that basil polysaccharide could significantly inhibit the invasiveness of SKOV3 cells in vitro in a dose-dependent manner, and that basil polysaccharides did not affect the expression of VEGF secretion in SKOV3 cells, nor did it affect the expression of VEGFR1 and VEGFR3 in SKOV3. However, the expression of mRNA and membrane surface protein in SKOV3 cells could be upregulated significantly by VEGFR2, and the expression of VEGFR2 in SKOV3 cells was significantly down-regulated by QRT-PCR and flow cytometry after successful infection with stable expression of lentivirus against VEGFR2/KDR shRNA. Further studies showed that basil polysaccharides significantly inhibited the invasion of SKOV3 cells expressing knockout VEGFR2. Conclusion basil polysaccharide, a traditional Chinese medicine extract, can significantly inhibit the invasion of ovarian cancer cell line SKOV3, which is mediated in part by up-regulating the expression of VEGFR2 in SKOV3 cells.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.31

【相似文獻】

相關期刊論文 前10條

1 郭文菁;楊美香;曲迅;鄭廣娟;張丹;孔北華;;羅勒多糖對樹突狀細胞表面分子表達的影響[J];中國免疫學雜志;2006年09期

2 郭文菁;馬秀明;楊美香;鄭廣娟;孔北華;曲迅;;羅勒多糖對卵巢癌生物學行為的影響及其機制研究[J];現代婦產科進展;2006年09期

3 趙慶華;趙豫;;羅勒多糖抗腫瘤轉移作用的機制研究[J];腫瘤藥學;2013年05期

4 曲迅,楊美香,鄭廣娟,郭文菁,周文,劉德山,張丹,張靜,趙麗霞,夏麗英;羅勒多糖對腫瘤轉移行為的影響[J];中國腫瘤生物治療雜志;2004年01期

5 鄒明瑾,郭文靜,楊美香,張丹,張靜,閆實,曲迅;表達譜芯片技術在羅勒多糖抗腫瘤機制研究中的應用[J];山東醫(yī)藥;2005年20期

6 劉向紅;楊美香;潘文英;鄭廣娟;張延;楊詠梅;鄧碧萍;夏麗英;曲迅;;羅勒多糖對人樹突細胞抗原攝取功能及共刺激分子表達的影響[J];中華微生物學和免疫學雜志;2007年09期

7 曲迅,鄭廣娟,劉德山,楊美香,馮進波,閆實;體外羅勒多糖抗人高轉移肺癌細胞侵襲轉移作用及機制探討[J];中國病理生理雜志;2005年07期

8 王婷;鄭廣娟;閆實;曲迅;;羅勒多糖抗卵巢癌侵襲轉移的體外實驗研究[J];中國病理生理雜志;2009年04期

9 楊京霞;黃銀生;屈長青;;羅勒多糖的提取及含量測定[J];阜陽師范學院學報(自然科學版);2012年02期

10 付偉;高文娟;張志勉;曲迅;趙玉霞;;羅勒多糖對卵巢癌SKOV3細胞VEGF受體表達的調控作用及對其侵襲能力的影響[J];山東大學學報(醫(yī)學版);2014年05期

相關會議論文 前1條

1 郭文菁;張彬彬;邵倩倩;高文娟;劉佳;王春娥;宋秉鳳;曲迅;;低氧環(huán)境下羅勒多糖對人乳腺癌MDA-231細胞株TIMPs表達的影響[A];第九屆全國腫瘤生物治療學術會議論文集[C];2006年

相關碩士學位論文 前6條

1 王婷;羅勒多糖抗卵巢癌細胞侵襲轉移的體外實驗研究[D];山東中醫(yī)藥大學;2008年

2 張彬彬;羅勒多糖對腫瘤細胞侵襲轉移活性的影響[D];昆明醫(yī)學院;2006年

3 付偉;羅勒多糖對卵巢癌SKOV3細胞血管內皮生長因子受體表達的調控作用及對其侵襲能力的影響[D];山東大學;2014年

4 張妲;羅勒多糖對人卵巢癌細胞SKOV3細胞骨架及SEMA4C表達影響的實驗研究[D];山東中醫(yī)藥大學;2013年

5 郭小顏;羅勒多糖對H22肝癌淋巴道轉移模型小鼠VEGF-C/VEGFR-3信號通路的影響[D];山東中醫(yī)藥大學;2013年

6 連松剛;羅勒多糖對人淋巴管內皮細胞（HLECs）VEGF-C/D-VEGFR-2/3信號通路及Bax和Bcl-2基因表達的影響[D];山東中醫(yī)藥大學;2014年

，

本文編號：1866793