索拉非尼对宫颈癌Hela细胞增殖影响的研究

发布时间：2018-12-18 15:36

【摘要】：目的：以宫颈癌Hela细胞为研究对象，采用分子靶向药物索拉非尼作用于宫颈癌Hela细胞，观察其对Hela细胞生长抑制作用的影响；研究索拉非尼联合顺铂、紫杉醇对Hela细胞生长抑制作用，观察索拉非尼对宫颈癌Hela细胞VEGFR-3和PDGFR-β表达的影响作用及其分子机制，为宫颈癌的生物治疗提供理论依据。方法:四甲基偶氮唑蓝（MTT）快速比色法检测索拉非尼作用于Hela细胞24h、48h、72h后细胞的增殖率，以及索拉非尼联合化疗药物（顺铂、紫杉醇）对Hela细胞增殖的抑制作用。采用Hoechst荧光染色观察药物作用48h后的细胞凋亡情况，Hela细胞的形态学变化。细胞免疫荧光法和Western blotting检测药物作用前后Hela细胞VEGFR-3和PDGFR-β表达情况的变化，并分析二者表达变化与Hela细胞增殖、凋亡变化的相关性。结果：索拉非尼对宫颈癌Hela细胞的增殖有明显抑制作用（P0.05）；随着药物浓度的增加，细胞增殖率不断下降，呈浓度依赖性。不同浓度的索拉非尼联合化疗药物（顺铂、紫杉醇）抑制Hela细胞的增殖，随着药物浓度的增加Hela细胞的增殖率下降，且比单用化疗药物抑制率高，索拉非尼联合化疗药物对Hela细胞的抑制具有协同作用；药物作用于Hela细胞后能观察到凋亡的Hela细胞，细胞核变小、固缩，荧光明显增强；正常Hela细胞中有VEGFR-3和PDGFR-β表达，，索拉非尼作用Hela细胞后，Hela细胞绿色荧光明显减弱，VEGFR-3和PDGFR-β表达明显降低,VEGFR-3和PDGFR-β表达变化与Hela细胞的增殖呈显著正相关，并且与Hela细胞的凋亡呈显著负相关。结论：（一）：索拉非尼抑制Hela细胞的增殖；（二）：索拉非尼联合化疗药物顺铂紫杉醇对宫颈癌Hela细胞的增殖抑制有协同作用；（三）：索拉非尼可能通过抑制Hela细胞中VEGFR-3和PDGFR-β的表达，发挥抑制宫颈癌细胞增殖的作用。
[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.
【学位授予单位】：河南科技大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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