【摘要】：目的探討研究vandetanib對人卵巢漿液性乳頭狀腺癌細胞株SKOV3和卵巢癌順鉑耐藥細胞株SKOV3/DDP的增殖抑制作用及其機制的探討。 方法采用MTT法檢測不同時間不同濃度的vandetanib作用SKOV3及SKOV3/DDP細胞的殺傷作用并計算半抑制濃度(IC50)；流式細胞術(shù)(FCM)檢測藥物對細胞凋亡及其周期分布變化；采用蛋白印跡法(Western blot)分析凋亡基因Bcl-2，Bax及反應(yīng)腫瘤細胞侵襲力的基因MMP-2的蛋白表達變化。 結(jié)果與對照組相比，vandetanib能明顯抑制SKOV3和SKOV3/DDP生長，呈時間-劑量依賴性，IC50值均呈明顯減小趨勢；32μmol/L和64μmol/L的vandetanib作用72h后，對SKOV3/DDP和SKOV3的抑制率分別達到56.74%和55.37%；64μmol/L vandetanib作用48h、32μmol/L和64μmol/L的vandetanib作用72h后，SKOV3/DDP的抑制率明顯高于SKOV3，差異有統(tǒng)計學(xué)意義（P0.05）。流式細胞術(shù)結(jié)果顯示隨時間及濃度的增加，兩種細胞凋亡明顯增加，G1期細胞增加，，S期和G2期細胞減少，差異有統(tǒng)計學(xué)意義(P0.05)。蛋白印跡法結(jié)果顯示，藥物作用于SKOV3敏感株Bcl-2、MMP-2表達減少，Bax表達無明顯變化；藥物作用于SKOV3/DDP耐藥株Bcl-2表達減少，Bax表達增加，MMP-2不表達。 結(jié)論Vandetanib對卵巢癌細胞株及順鉑耐藥株的生長有明顯的呈時間及濃度依賴性的抑制作用，并且達到一定濃度后，對耐藥株SKOV3/DDP細胞的抑制作用較敏感株SKOV3明顯增強；vandetanib抑制卵巢癌的生長可能與細胞周期的G0/G1期阻滯有關(guān)；通過降低Bcl-2/Bax比值可能是vandetanib誘導(dǎo)細胞凋亡的一機制；vandetanib可能通過下調(diào)MMP-2來抑制原發(fā)卵巢癌細胞的侵襲轉(zhuǎn)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.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.31

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