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兩種方法建立的睪丸癌順鉑耐藥細(xì)胞株的比較

發(fā)布時(shí)間:2018-06-22 09:51

  本文選題:睪丸癌 + 順鉑 ; 參考:《南方醫(yī)科大學(xué)學(xué)報(bào)》2015年12期


【摘要】:目的濃度遞增法與大劑量沖擊法分別誘導(dǎo)兩種小鼠睪丸癌I-10順鉑耐藥細(xì)胞株,比較兩種細(xì)胞株之間形態(tài)差異,并檢測(cè)耐藥相關(guān)蛋白MDR1及P-gp的表達(dá)水平。方法采用濃度遞增法誘導(dǎo)耐藥細(xì)胞株I-10/DDPi,大劑量沖擊法的誘導(dǎo)耐藥細(xì)胞株I-10/DDPh;顯微鏡下觀察細(xì)胞形態(tài)改變;MTT法分別檢測(cè)耐藥指數(shù),繪制生長(zhǎng)曲線;Western blot檢測(cè)耐藥相關(guān)蛋白MDR1及Pgp的表達(dá);Transwell法檢測(cè)細(xì)胞侵襲能力的變化。結(jié)果正常睪丸癌I-10細(xì)胞及兩種方法建立的相應(yīng)耐藥株均為貼壁生長(zhǎng),形態(tài)飽滿,大體呈梭形,上皮樣,細(xì)胞大小大致相同;I-10/DDP細(xì)胞大小不同,形態(tài)不規(guī)則,細(xì)胞表面有細(xì)長(zhǎng)微絨毛類似觸角,分散排列。耐藥株I-10/DDPi及I-10/DDPh對(duì)順鉑的耐藥指數(shù)分別為3.924和3.099,耐藥細(xì)胞系的倍增時(shí)間較正常I-10細(xì)胞延長(zhǎng)。耐藥細(xì)胞中耐藥相關(guān)蛋白MDR1及P-gp表達(dá)顯著高于正常I-10細(xì)胞,其中I-10/DDPi增加更為明顯。耐藥細(xì)胞株侵襲力增強(qiáng),其中I-10/DDPh增加更為明顯。結(jié)論采用兩種方法均可成功建立的耐順鉑睪丸癌細(xì)胞株,且大劑量沖擊法建立的耐順鉑細(xì)胞株I-10/DDPh更接近于臨床中的腫瘤耐藥細(xì)胞。
[Abstract]:Objective to detect the expression of MDR1 and P-gp in mouse testicular carcinoma cells induced by increasing concentration and high dose shock respectively, and to compare the morphological differences between the two cell lines. Methods the drug resistant cell line I-10 / DDPiand the I-10 / DDPh-induced drug resistant cell line were induced by increasing concentration, and the cell morphologic changes were observed under microscope and the drug resistance index was detected by MTT method. The expression of MDR1 and PGP was detected by Western blot. Results the normal testicular carcinoma I-10 cells and the corresponding drug-resistant strains were all adherent to the wall, full in shape, generally fusiform, epithelioid, similar in size and irregular in size of I-10 / DDP cells. Cell surface has slender microvilli similar to the antennae, scattered arrangement. The resistance index of resistant cell lines I-10 / DDPi and I-10 / DDPh to cisplatin were 3.924 and 3.099, respectively. The doubling time of resistant cell lines was longer than that of normal I-10 cells. The expression of MDR1 and P-gp in drug-resistant cells was significantly higher than that in normal I-10 cells, especially in I-10 / DDPi cells. The invasiveness of drug resistant cell line was increased, especially I 10 / DDPh. Conclusion Cisplatin-resistant testicular cancer cell line can be successfully established by both methods, and the cisplatin resistant cell line I-10 / DDPh established by high-dose impulse method is closer to clinical tumor resistant cell line.
【作者單位】: 蚌埠醫(yī)學(xué)院藥學(xué)系藥理學(xué)教研室;
【基金】:安徽高校自然科學(xué)研究項(xiàng)目(KJ2015A180) 蚌埠醫(yī)學(xué)院校級(jí)自然科學(xué)基金重點(diǎn)項(xiàng)目(BYKY1407ZD);蚌埠醫(yī)學(xué)院研究生科研創(chuàng)新計(jì)劃項(xiàng)目(Byycxz1420,Byycx1553) 高校優(yōu)秀青年人才支持計(jì)劃重點(diǎn)項(xiàng)目(gxyq ZD2016158)
【分類號(hào)】:R737.21

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