發(fā)布時(shí)間：2018-03-20 03:37

  本文選題：hepaCAM　切入點(diǎn)：吡柔比星　出處：《重慶醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：第一部分hepaCAM基因聯(lián)合吡柔比星增強(qiáng)對(duì)膀胱癌細(xì)胞BIU-87增殖的研究 目的：研究肝細(xì)胞粘附分子(hepatocyte cell adhesion molecule，hepaCAM)聯(lián)合吡柔比星（pirarubicin, THP）對(duì)膀胱癌細(xì)胞BIU-87增殖的影響。 方法：用腺病毒向膀胱癌BIU-87導(dǎo)入hepaCAM基因，加吡柔比星處理后，采用實(shí)時(shí)熒光定量PCR檢測(cè)pAdH5-hepaCAM處理組，吡柔比星處理組，吡柔比星聯(lián)合pAdH5-hepaCAM處理組和未感染組的膀胱癌BIU-87細(xì)胞中相關(guān)基因cyclinD1、CDK2的mRNA表達(dá)水平，同時(shí)MTT測(cè)各處理組細(xì)胞抑制率及FCM測(cè)各處理組細(xì)胞周期變化。 結(jié)果：實(shí)時(shí)熒光定量PCR結(jié)果顯示：吡柔比星聯(lián)合pAdH5-hepaCAM處理組，較單獨(dú)處理組，cyclinD1、CDK2的mRNA水平明顯降低（P0.05）； MTT檢測(cè)的各處理組中聯(lián)合處理組細(xì)胞增殖明顯受到抑制；FCM檢測(cè)證實(shí)，hepaCAM基因和吡柔比星聯(lián)合處理組細(xì)胞G0/G1期較空白組和hepaCAM基因、吡柔比星單獨(dú)處理組相比，明顯升高（P0.05）。 結(jié)論： hepaCAM基因感染膀胱癌細(xì)胞BIU-87，再給予吡柔比星藥物處理后，細(xì)胞增殖受到抑制和細(xì)胞周期出現(xiàn)了明顯的G0/G1阻滯，因此，hepaCAM基因聯(lián)合吡柔比星可以增強(qiáng)對(duì)膀胱癌細(xì)胞增殖作用。 第二部分hepaCAM基因?qū)Π螂装┠[瘤生長(zhǎng)體內(nèi)實(shí)驗(yàn)研究 目的：研究hepaCAM基因?qū)Π螂装〣IU-87細(xì)胞株裸鼠移植腫瘤生長(zhǎng)情況的影響。 方法：將9只裸鼠隨機(jī)為空載組和hepaCAM組，于裸鼠右下肢背外側(cè)皮下接種BIU-87細(xì)胞l×107/只，接種完畢后放SPF環(huán)境培養(yǎng)，，每天定時(shí)觀察成瘤情況并記錄瘤體體積和裸鼠重量。三周后處死老鼠，取瘤體比較兩組裸鼠的成瘤大小，同時(shí)取瘤組織做免疫組化檢測(cè)hepaCAM蛋白的表達(dá)。 結(jié)果：空白組與感染組相比較，瘤體積明顯較大（p0.05），免疫組化顯示空白組hepaCAM蛋白表達(dá)比感染組明顯減弱。 結(jié)論：以腺病毒為載體，體內(nèi)實(shí)驗(yàn)進(jìn)一步證實(shí)hepaCAM基因能明顯抑制膀胱腫瘤的生長(zhǎng)。因此，以腺病毒為載體的hepaCAM基因有可能成為一種有效的基因治療方法。
[Abstract]:A study on the effect of hepaCAM gene combined with pirarubicin on the proliferation of bladder cancer cell line BIU-87. Aim: to study the effect of hepatocyte cell adhesion molecule (hepatocyte cell adhesion molecule) combined with pirarubicin (THP1) on BIU-87 proliferation of bladder cancer cells. Methods: hepaCAM gene was introduced into bladder cancer BIU-87 by adenovirus. After treated with pirarubicin, real-time fluorescence quantitative PCR was used to detect pAdH5-hepaCAM treatment group and pirarubicin treatment group. The mRNA expression of cyclin D1 CDK2 was detected in bladder cancer BIU-87 cells treated with pirarubicin combined with pAdH5-hepaCAM, and the cell cycle changes of each treatment group were measured by MTT and FCM. Results: the results of real-time fluorescence quantitative PCR showed that Pirarubicin combined with pAdH5-hepaCAM treatment group, The level of mRNA in cyclin D1 + CDK2 group was significantly lower than that in the control group (P 0.05), and the cell proliferation in the combined treatment group was significantly inhibited by MTT. The results of FCM showed that the G0 / P G 1 phase of the treated group was significantly lower than that of the control group and the hepaCAM gene of the combined treatment group was higher than that of the control group, and that of the combined treatment group was significantly lower than that of the control group and the hepaCAM gene group. Compared with the group treated with pirarubicin alone, it was significantly higher than that of the control group (P 0.05). Conclusion: after hepaCAM gene was infected with BIU-87 and then treated with pirarubicin, cell proliferation was inhibited and cell cycle was significantly blocked by G _ 0 / G _ 1. Therefore, hepaCAM gene combined with pirarubicin could enhance the proliferation of bladder cancer cells. The second part: an in vivo study on the effect of hepaCAM gene on the growth of bladder cancer. Aim: to study the effect of hepaCAM gene on the growth of bladder cancer BIU-87 cell line in nude mice. Methods: nine nude mice were randomly divided into no-load group and hepaCAM group. BIU-87 cells were inoculated subcutaneously on the dorsolateral side of right lower extremity of nude mice. After inoculation, BIU-87 cells were cultured in SPF environment. The tumor size and weight of nude mice were recorded three weeks later. The tumor size of the two groups was compared and the expression of hepaCAM protein was detected by immunohistochemistry. Results: compared with the infected group, the tumor volume of the blank group was significantly larger than that of the infected group, and the expression of hepaCAM protein in the blank group was significantly lower than that in the infected group. Conclusion: using adenovirus as vector, it is further confirmed that hepaCAM gene can inhibit the growth of bladder tumor in vivo. Therefore, hepaCAM gene with adenovirus as vector may become an effective gene therapy method.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.14

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本文編號(hào)：1637368