發(fā)布時(shí)間：2018-12-18 15:36

【摘要】：目的：以宮頸癌Hela細(xì)胞為研究對(duì)象，采用分子靶向藥物索拉非尼作用于宮頸癌Hela細(xì)胞，觀察其對(duì)Hela細(xì)胞生長(zhǎng)抑制作用的影響；研究索拉非尼聯(lián)合順鉑、紫杉醇對(duì)Hela細(xì)胞生長(zhǎng)抑制作用，觀察索拉非尼對(duì)宮頸癌Hela細(xì)胞VEGFR-3和PDGFR-β表達(dá)的影響作用及其分子機(jī)制，為宮頸癌的生物治療提供理論依據(jù)。 方法:四甲基偶氮唑藍(lán)（MTT）快速比色法檢測(cè)索拉非尼作用于Hela細(xì)胞24h、48h、72h后細(xì)胞的增殖率，以及索拉非尼聯(lián)合化療藥物（順鉑、紫杉醇）對(duì)Hela細(xì)胞增殖的抑制作用。采用Hoechst熒光染色觀察藥物作用48h后的細(xì)胞凋亡情況，Hela細(xì)胞的形態(tài)學(xué)變化。細(xì)胞免疫熒光法和Western blotting檢測(cè)藥物作用前后Hela細(xì)胞VEGFR-3和PDGFR-β表達(dá)情況的變化，并分析二者表達(dá)變化與Hela細(xì)胞增殖、凋亡變化的相關(guān)性。 結(jié)果：索拉非尼對(duì)宮頸癌Hela細(xì)胞的增殖有明顯抑制作用（P0.05）；隨著藥物濃度的增加，細(xì)胞增殖率不斷下降，呈濃度依賴性。不同濃度的索拉非尼聯(lián)合化療藥物（順鉑、紫杉醇）抑制Hela細(xì)胞的增殖，隨著藥物濃度的增加Hela細(xì)胞的增殖率下降，且比單用化療藥物抑制率高，索拉非尼聯(lián)合化療藥物對(duì)Hela細(xì)胞的抑制具有協(xié)同作用；藥物作用于Hela細(xì)胞后能觀察到凋亡的Hela細(xì)胞，細(xì)胞核變小、固縮，熒光明顯增強(qiáng)；正常Hela細(xì)胞中有VEGFR-3和PDGFR-β表達(dá)，，索拉非尼作用Hela細(xì)胞后，Hela細(xì)胞綠色熒光明顯減弱，VEGFR-3和PDGFR-β表達(dá)明顯降低,VEGFR-3和PDGFR-β表達(dá)變化與Hela細(xì)胞的增殖呈顯著正相關(guān)，并且與Hela細(xì)胞的凋亡呈顯著負(fù)相關(guān)。 結(jié)論：（一）：索拉非尼抑制Hela細(xì)胞的增殖；（二）：索拉非尼聯(lián)合化療藥物順鉑紫杉醇對(duì)宮頸癌Hela細(xì)胞的增殖抑制有協(xié)同作用；（三）：索拉非尼可能通過(guò)抑制Hela細(xì)胞中VEGFR-3和PDGFR-β的表達(dá)，發(fā)揮抑制宮頸癌細(xì)胞增殖的作用。
[Abstract]:Objective: to observe the effect of Solafenib, a molecular targeted drug, on the growth inhibition of cervical cancer Hela cells in Hela cells. To study the inhibitory effect of Solafenil combined with cisplatin and paclitaxel on the growth of Hela cells, and to observe the effect of Solafenil on the expression of VEGFR-3 and PDGFR- 尾 in Hela cells of cervical cancer and its molecular mechanism, and to provide a theoretical basis for the biotherapy of cervical cancer. Methods: rapid colorimetric assay of tetramethylazolium blue (MTT) was used to detect the proliferation rate of Hela cells treated with solafenil for 24 h or 48 h or 72 h, and the inhibitory effect of Solafenil combined with chemotherapeutic drugs (cisplatin, paclitaxel) on the proliferation of Hela cells. Apoptosis and morphological changes of Hela cells were observed by Hoechst fluorescence staining after 48 h treatment. The changes of VEGFR-3 and PDGFR- 尾 expression in Hela cells before and after drug treatment were detected by cell immunofluorescence assay and Western blotting. The relationship between the changes of VEGFR-3 and PDGFR- 尾 expression and the proliferation and apoptosis of Hela cells was analyzed. Results: Solafenib significantly inhibited the proliferation of cervical cancer Hela cells (P0.05), and with the increase of drug concentration, cell proliferation rate decreased in a concentration-dependent manner. Different concentrations of Solafenil combined with chemotherapy (cisplatin, paclitaxel) inhibited the proliferation of Hela cells. With the increase of drug concentration, the proliferation rate of Hela cells decreased, and the inhibition rate was higher than that of chemotherapy drugs alone. Solafenib combined with chemotherapeutic drugs had synergistic effect on the inhibition of Hela cells. The apoptotic Hela cells were observed after treatment with the drug. The nuclei became smaller, pyknosis and fluorescence increased significantly. The expression of VEGFR-3 and PDGFR- 尾 was observed in normal Hela cells. After treated with Solafenil, the green fluorescence of Hela cells decreased, and the expression of VEGFR-3 and PDGFR- 尾 decreased significantly. The expression of VEGFR-3 and PDGFR- 尾 was positively correlated with the proliferation of Hela cells and negatively correlated with the apoptosis of Hela cells. Conclusion: (1) Solafenib inhibits the proliferation of Hela cells, (2) Solafenib combined with cisplatin paclitaxel has synergistic effect on the proliferation of cervical cancer Hela cells. (3) Solafenib may inhibit the proliferation of cervical cancer cells by inhibiting the expression of VEGFR-3 and PDGFR- 尾 in Hela cells.
【學(xué)位授予單位】：河南科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R737.33

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相關(guān)期刊論文 前1條

1 曹澤毅;子宮頸癌治療的變遷和思考[J];中華婦產(chǎn)科雜志;2004年03期

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