LRIG1在卵巢漿液性囊腺癌細胞對依托泊苷敏感性中的作用研究
發(fā)布時間:2018-02-02 16:09
本文關(guān)鍵詞: 卵巢癌 耐藥 亮氨酸重復序列免疫球蛋白樣結(jié)構(gòu)域基因- 出處:《中國現(xiàn)代醫(yī)學雜志》2017年11期 論文類型:期刊論文
【摘要】:目的探討亮氨酸重復序列免疫球蛋白樣結(jié)構(gòu)域基因-1(LRIG1)在卵巢漿液性囊腺癌(SKOV3)細胞株中對依托泊苷敏感性的可能機制。方法噻唑藍比色法(MMT)檢測不同濃度VP16干預下48 h的SKOV3細胞組、siRNA LRIG1轉(zhuǎn)染SKOV3細胞組、SKOV3/VP16細胞組和siRNA LRIG1轉(zhuǎn)染SKOV3/VP16細胞組的增殖。平板克隆檢測細胞增殖,流式檢測細胞凋亡情況,實時熒光定量聚合酶鏈反應(yīng)(q RT-PCR)檢測LRIG1mRNA表達。結(jié)果半抑制濃度(IC50)分別為62.90、115.49、156.50和195.42μg/L,siRNA LRIG1轉(zhuǎn)染SKOV3、SKOV3/VP16和siRNA LRIG1轉(zhuǎn)染SKOV3/VP16的耐藥指數(shù)分別為1.8、2.5和3.1。SKOV3、siRNA LRIG1轉(zhuǎn)染SKOV3、SKOV3/VP16和siRNA LRIG1轉(zhuǎn)染SKOV3/VP16細胞組的克隆率(F=39.338,P=0.000),細胞的LRIG1 mRNA(F=63.095,P=0.000)和凋亡細胞(F=230.046,P=0.000)明顯不同,與SKOV3比較,siRNA LRIG1轉(zhuǎn)染SKOV3、SKOV3/VP16和siRNA LRIG1轉(zhuǎn)染SKOV3/VP16細胞的克隆率增加(t=0.026、0.0710和0.125,P=0.042、0.000和0.000),細胞的LRIG1 mRNA降低(t=0.130、0.525和0.825,均P=0.000),凋亡細胞減少(t=12.350、35.506和44.412,均P=0.000),與siRNA LRIG1轉(zhuǎn)染SKOV3比較,SKOV3/VP16和siRNA LRIG1轉(zhuǎn)染SKOV3/VP16細胞的克隆率增加(t=0.044和0.099,P=0.001和0.000),LRIG1 mRNA降低(t=0.395和0.695,均P=0.000),凋亡細胞減少(t=23.156和32063,均P0.05),與SKOV3/VP16比較,siRNA LRIG1轉(zhuǎn)染SKOV3/VP16細胞的克隆率增加(t=0.055,P=0.000),細胞的LRIG1 mRNA降低(t=0.300,P=0.000),凋亡細胞減少(t=8.906,P=0.000)。結(jié)論 LRIG1 mRNA影響SKOV3細胞對藥物的敏感性,沉默LRIG1的耐藥細胞可抑制SKOV3細胞凋亡。
[Abstract]:Objective to investigate the role of leucine repeat immunoglobulin-like domain gene (LRIG1) in ovarian serous cystadenocarcinoma (SKOV3). Methods the possible mechanism of sensitivity to etoposide in cell lines. Methods the SKOV3 cells treated with different concentrations of VP16 for 48 h were detected by thiazolyl blue colorimetric assay. SiRNA LRIG1 was transfected into SKOV3 cells. The proliferation of SKOV3/VP16 cells and SKOV3/VP16 cells transfected with siRNA LRIG1 was observed. The proliferation of SKOV3/VP16 cells was detected by plate clone and apoptosis was detected by flow cytometry. The expression of LRIG1mRNA was detected by real-time fluorescence quantitative polymerase chain reaction (Q RT-PCR). Results the semi-inhibitory concentration of IC50 was 62.90 ~ 115.49, respectively. 156.50 and 195.42 渭 g / L siRNA LRIG1 were transfected into SKOV3. The drug resistance index of SKOV3/VP16 transfected with SKOV3/VP16 and siRNA LRIG1 was 1.82.5 and 3.1.SKOV3, respectively. The clone rate of SKOV3/VP16 cells transfected with siRNA LRIG1 and SKOV3 / SKOV3 / VP16 and siRNA LRIG1 was 39.338. The LRIG1 mRNAs of the cells were significantly different from that of the apoptotic cells. Compared with SKOV3, siRNA LRIG1 was transfected into SKOV3. The clone rates of SKOV3/VP16 cells transfected with SKOV3/VP16 and siRNA LRIG1 were increased by 0.026, 0.0710 and 0.125, respectively. The LRIG1 mRNA of the cells decreased from 0.130 to 0.825, both P0. 000). The number of apoptotic cells decreased from 12.350 to 35.506 and 44.412, respectively, compared with that of siRNA LRIG1 transfected SKOV3. The clone rates of SKOV3/VP16 cells transfected with SKOV3/VP16 and siRNA LRIG1 were increased by 0.044 and 0.099 respectively. P0. 001 and 0. 000 LRIG1 mRNA were decreased by 0. 395 and 0. 695, respectively (P 0. 000). Apoptotic cells were decreased by 23.156 and 32063, both P0.05, compared with SKOV3/VP16. The clone rate of SKOV3/VP16 cells transfected with siRNA LRIG1 was increased by 0.055 P0. 000). The LRIG1 mRNA of the cells was decreased by 0.300, and the apoptotic cells were decreased by 8.906. Conclusion LRIG1 mRNA affects the drug sensitivity of SKOV3 cells and silencing LRIG1 resistant cells can inhibit the apoptosis of SKOV3 cells.
【作者單位】: 桂林醫(yī)學院第二附屬醫(yī)院婦科;桂林醫(yī)學院附屬醫(yī)院婦科;桂林醫(yī)學院第二附屬醫(yī)院病理科;桂林醫(yī)學院附屬醫(yī)院統(tǒng)計室;
【基金】:桂林市科學研究與技術(shù)開發(fā)計劃項目(No:20130120-3)
【分類號】:R737.31
【正文快照】: 當今,卵巢癌的發(fā)生率已經(jīng)呈現(xiàn)出不斷上升的1材料與方法趨勢,成為危害女性的重要殺手,發(fā)病率僅次于子1.1材料和試劑宮頸癌和子宮體癌而列居第3位,卵巢腫瘤中的卵卵巢漿液性囊腺癌(SKOV3)細胞株(北京大學巢上皮癌的死亡為女性各種腫瘤死亡之首,可見卵人民醫(yī)院),依托泊苷(VP16)(,
本文編號:1484923
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