Vandetanib对人卵巢癌细胞抑制作用及其机制的探讨
发布时间：2018-11-14 15:54
【摘要】：目的探讨研究vandetanib对人卵巢浆液性乳头状腺癌细胞株SKOV3和卵巢癌顺铂耐药细胞株SKOV3/DDP的增殖抑制作用及其机制的探讨。 方法采用MTT法检测不同时间不同浓度的vandetanib作用SKOV3及SKOV3/DDP细胞的杀伤作用并计算半抑制浓度(IC50)；流式细胞术(FCM)检测药物对细胞凋亡及其周期分布变化；采用蛋白印迹法(Western blot)分析凋亡基因Bcl-2，Bax及反应肿瘤细胞侵袭力的基因MMP-2的蛋白表达变化。 结果与对照组相比，vandetanib能明显抑制SKOV3和SKOV3/DDP生长，呈时间-剂量依赖性，IC50值均呈明显减小趋势；32μmol/L和64μmol/L的vandetanib作用72h后，对SKOV3/DDP和SKOV3的抑制率分别达到56.74%和55.37%；64μmol/L vandetanib作用48h、32μmol/L和64μmol/L的vandetanib作用72h后，SKOV3/DDP的抑制率明显高于SKOV3，差异有统计学意义（P0.05）。流式细胞术结果显示随时间及浓度的增加，两种细胞凋亡明显增加，G1期细胞增加，，S期和G2期细胞减少，差异有统计学意义(P0.05)。蛋白印迹法结果显示，药物作用于SKOV3敏感株Bcl-2、MMP-2表达减少，Bax表达无明显变化；药物作用于SKOV3/DDP耐药株Bcl-2表达减少，Bax表达增加，MMP-2不表达。 结论Vandetanib对卵巢癌细胞株及顺铂耐药株的生长有明显的呈时间及浓度依赖性的抑制作用，并且达到一定浓度后，对耐药株SKOV3/DDP细胞的抑制作用较敏感株SKOV3明显增强；vandetanib抑制卵巢癌的生长可能与细胞周期的G0/G1期阻滞有关；通过降低Bcl-2/Bax比值可能是vandetanib诱导细胞凋亡的一机制；vandetanib可能通过下调MMP-2来抑制原发卵巢癌细胞的侵袭转移。
[Abstract]:Objective to investigate the inhibitory effect of vandetanib on the proliferation of human ovarian serous papillary adenocarcinoma cell line SKOV3 and cisplatin resistant ovarian cancer cell line SKOV3/DDP and its mechanism. Methods MTT assay was used to detect the cytotoxicity of SKOV3 and SKOV3/DDP cells at different time and concentration of vandetanib, and the semi-inhibitory concentration (IC50) was calculated, and the changes of cell apoptosis and cell cycle distribution were detected by flow cytometry (FCM) (FCM). The protein expression of apoptotic gene Bcl-2,Bax and tumor cell invasiveness gene MMP-2 was analyzed by Western blot (Western blot). Results compared with the control group, vandetanib inhibited the growth of SKOV3 and SKOV3/DDP in a time-dose dependent manner, and the IC50 value decreased significantly. After treated with 32 渭 mol/L and 64 渭 mol/L vandetanib for 72 h, the inhibition rates of SKOV3/DDP and SKOV3 were 56.74% and 55.37%, respectively. After treated with 64 渭 mol/L vandetanib for 48 h, 32 渭 mol/L and 64 渭 mol/L vandetanib for 72 h, the inhibition rate of SKOV3/DDP was significantly higher than that of SKOV3, (P0.05). The results of flow cytometry showed that with the increase of time and concentration, apoptosis of the two types of cells increased significantly, G1 phase cells increased, S phase and G2 phase cells decreased, the difference was statistically significant (P0.05). The results of Western blotting showed that the expression of Bcl-2,MMP-2 decreased, but the expression of Bax did not change in the sensitive SKOV3 strain, but the expression of Bcl-2 decreased, the expression of Bax increased, but the expression of MMP-2 did not change. Conclusion Vandetanib can inhibit the growth of ovarian cancer cell line and cisplatin resistant cell line in a time-and concentration-dependent manner, and the inhibitory effect of Vandetanib on SKOV3/DDP cell line is stronger than that on the sensitive cell line SKOV3. The inhibition of the growth of ovarian cancer by vandetanib may be related to the arrest of G0/G1 phase of cell cycle, and the decrease of Bcl-2/Bax ratio may be a mechanism of apoptosis induced by vandetanib. Vandetanib may inhibit the invasion and metastasis of primary ovarian cancer cells by down-regulating MMP-2.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31
本文编号：2331617
[Abstract]:Objective to investigate the inhibitory effect of vandetanib on the proliferation of human ovarian serous papillary adenocarcinoma cell line SKOV3 and cisplatin resistant ovarian cancer cell line SKOV3/DDP and its mechanism. Methods MTT assay was used to detect the cytotoxicity of SKOV3 and SKOV3/DDP cells at different time and concentration of vandetanib, and the semi-inhibitory concentration (IC50) was calculated, and the changes of cell apoptosis and cell cycle distribution were detected by flow cytometry (FCM) (FCM). The protein expression of apoptotic gene Bcl-2,Bax and tumor cell invasiveness gene MMP-2 was analyzed by Western blot (Western blot). Results compared with the control group, vandetanib inhibited the growth of SKOV3 and SKOV3/DDP in a time-dose dependent manner, and the IC50 value decreased significantly. After treated with 32 渭 mol/L and 64 渭 mol/L vandetanib for 72 h, the inhibition rates of SKOV3/DDP and SKOV3 were 56.74% and 55.37%, respectively. After treated with 64 渭 mol/L vandetanib for 48 h, 32 渭 mol/L and 64 渭 mol/L vandetanib for 72 h, the inhibition rate of SKOV3/DDP was significantly higher than that of SKOV3, (P0.05). The results of flow cytometry showed that with the increase of time and concentration, apoptosis of the two types of cells increased significantly, G1 phase cells increased, S phase and G2 phase cells decreased, the difference was statistically significant (P0.05). The results of Western blotting showed that the expression of Bcl-2,MMP-2 decreased, but the expression of Bax did not change in the sensitive SKOV3 strain, but the expression of Bcl-2 decreased, the expression of Bax increased, but the expression of MMP-2 did not change. Conclusion Vandetanib can inhibit the growth of ovarian cancer cell line and cisplatin resistant cell line in a time-and concentration-dependent manner, and the inhibitory effect of Vandetanib on SKOV3/DDP cell line is stronger than that on the sensitive cell line SKOV3. The inhibition of the growth of ovarian cancer by vandetanib may be related to the arrest of G0/G1 phase of cell cycle, and the decrease of Bcl-2/Bax ratio may be a mechanism of apoptosis induced by vandetanib. Vandetanib may inhibit the invasion and metastasis of primary ovarian cancer cells by down-regulating MMP-2.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31
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