本文關鍵詞： 白細胞介素-24 卵巢癌細胞 生長抑制 細胞凋亡 細胞周期 多基因遺傳表達分析　出處：《遵義醫(yī)學院》2014年碩士論文　論文類型：學位論文

【摘要】：目的： ①探討rhIL-24蛋白與順鉑聯(lián)合體外對人卵巢癌SKOV3細胞、人卵巢癌耐順鉑株SKOV3/ddp細胞的抑制效應； ②探討rhIL-24蛋白聯(lián)合順鉑體外誘導人卵巢癌SKOV3細胞、人卵巢癌耐順鉑株SKOV3/ddp細胞凋亡效應、細胞周期及凋亡基因的變化； ③探討腺病毒介導的白細胞介素-24基因與順鉑聯(lián)合體外誘導人卵巢癌SKOV3細胞的抑制效應及細胞凋亡情況變化。 方法： ①用rhIL-24、DDP及rhIL-24+DDP作用于SKOV3細胞、SKOV3/ddp細胞，用Ad-hIL-24、DDP及Ad-hIL-24+DDP作用于SKOV3細胞抑制細胞生長； ②應用CCK-8法檢測SKOV3細胞、SKOV3/ddp細胞增殖情況，計算抑制率； ③應用流式細胞儀檢測SKOV3細胞、SKOV3/ddp細胞凋亡及細胞周期的變化； ④應用多基因遺傳表達分析SKOV3細胞、SKOV3/ddp細胞凋亡多基因的變化。 結果： ①SKOV3細胞、SKOV3/ddp細胞在適宜條件下培養(yǎng)于第3天進入對數(shù)生長期； ②rhIL-24蛋白能抑制SKOV3細胞、SKOV3/ddp細胞生長，48小時的抑制率分別為30.7%、21.8%，對正常細胞無毒性作用；聯(lián)合DDP組抑制率分別為35.3%、32.6%，優(yōu)于單獨用藥組，，rhIL-24蛋白具有化療增敏效應； ③流式細胞儀檢測rhIL-24、DDP和rhIL-24+DDP各組凋亡率分別與陰性對照組比較，差異有顯著性意義；細胞周期檢測中，rhIL-24作用SKOV3細胞、SKOV3/ddp細胞，分別引起G2期阻滯，G2期及S期阻滯。 ④rhIL-24蛋白對于SKOV3細胞、SKOV3/ddp細胞具有上調(diào)Bax，下調(diào)Bcl-2，下調(diào)survivin，激活caspase3,上調(diào)抑癌基因Rb等凋亡相關基因，聯(lián)合DDP組中作用效果更加顯著。 ⑤Ad-hIL-24能抑制SKOV3細胞的生長，且聯(lián)合用藥之后對細胞生長抑制率明顯優(yōu)于單獨用藥組，Ad-hIL-24具有化療增敏效應。 結論： ①rhIL-24蛋白及Ad-hIL-24均能抑制SKOV3細胞、SKOV3/ddp細胞的體外生長，分別聯(lián)合DDP后效果優(yōu)于單獨用藥組，具有化療增敏效應，同時能誘導SKOV3細胞、SKOV3/ddp細胞凋亡。 ②rhIL-24蛋白可影響SKOV3細胞、SKOV3/ddp細胞的細胞周期，且能引起細胞凋亡相關基因的變化。
[Abstract]:Objective: 1 to investigate the inhibitory effect of rhIL-24 protein combined with cisplatin on human ovarian cancer SKOV3 cells and cisplatin resistant SKOV3/ddp cells. (2) to investigate the effect of rhIL-24 protein combined with cisplatin on inducing apoptosis of human ovarian cancer SKOV3 cells in vitro, and the changes of cell cycle and apoptotic genes in cisplatin-resistant SKOV3/ddp cells. 3 to investigate the inhibitory effect of adenovirus-mediated interleukin-24 gene and cisplatin combined with cisplatin on human ovarian cancer SKOV3 cells and the changes of apoptosis. Methods: 1 rhIL-24 DDP and rhIL-24 DDP were used to treat SKOV3 cells with SKOV3 / DDP cells and Ad-hIL-24. DDP and Ad-hIL-24 DDP inhibited the growth of SKOV3 cells. (2) the proliferation of SKOV3 / DDP cells in SKOV3 cells was detected by CCK-8 assay, and the inhibitory rate was calculated. 3The apoptosis and cell cycle of SKOV3 cell line SKOV3 / DDP were detected by flow cytometry. 4 Polygene expression was used to analyze the change of apoptotic polygene in SKOV3 cell line SKOV3 / DDP. Results: (1) SKOV3 cells entered logarithmic growth stage on the 3rd day. 2the inhibitory rates of rhIL-24 protein on the growth of SKOV3 / DDP cells in SKOV3 cells were 30.7 and 21.8h, respectively, and there was no toxic effect on normal cells. The inhibitory rates of rhIL-24 protein in combination with DDP group were 35.3and 32.6, respectively, which were superior to those of rhIL-24 protein alone in chemosensitization. 3The apoptotic rate of DDP and rhIL-24 DDP was detected by flow cytometry, which was significantly higher than that of negative control group. RhIL-24 induced the arrest of SKOV3 / DDP cells in SKOV3 cells in G2 phase and S-phase arrest, respectively. 4 rhIL-24 protein up-regulated Bax-2, down-regulated Bcl-2, down-regulated survivin and activated caspase3 in SKOV3 cell line SKOV3 / DDP. Apoptosis related genes such as RB were upregulated in combination with DDP. 5 Ad-hIL-24 could inhibit the growth of SKOV3 cells, and the inhibitory rate of Ad-hIL-24 was significantly higher than that of Ad-hIL-24 alone. Conclusion: 1rhIL-24 protein and Ad-hIL-24 could inhibit the growth of SKOV3 / DDP cells in SKOV3 cells in vitro, and the effect after combined with DDP was better than that in the control group. It has chemosensitization effect and induces apoptosis of SKOV3 / DDP cells in SKOV3 cells. 2rhIL-24 protein can affect the cell cycle of SKOV3 / DDP cells in SKOV3 cells and induce the changes of apoptosis-related genes.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.31

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