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醫(yī)學免費論文:雷帕霉素對小鼠H22肝癌細胞生長增殖的影響
【摘要】 目的 研究雷帕霉素(rapamycin, RPM)對H22肝癌細胞生長增殖的影響。方法 體外培養(yǎng)小鼠H22肝細胞癌株，分別與RPM、CsA、FK506和5Fu共同孵育48h，進行MTT實驗。流式細胞儀測定各組H22細胞的細胞周期變化，ELISA法測定各組上清液中VEGF含量。體內(nèi)實驗建立H22肝細胞癌皮下移植瘤模型，同時完成C57BL/6→BALB/c小鼠異體皮膚移植模型。給予RPM、CsA、FK506和5Fu灌胃，觀察皮片存活情況。獲取實驗小鼠血清及腫瘤組織。計算各組腫瘤體積，通過CD34免疫組化染色測定各組腫瘤組織的微血管密度(MVD)。結果 劑量為0.01、0.1、1mg/L的RPM對對數(shù)生長的H22肝細胞具有細胞毒性，抑制H22小鼠肝癌細胞增殖，培養(yǎng)上清液中的VEGF濃度較對照組顯著降低(P<0.05)，細胞周期分析顯示S期細胞數(shù)較其他免疫抑制劑組顯著減少(P<0.05)。體內(nèi)實驗顯示給予1.5mg/(kg·d)和4.5mg/(kg·d)的RPM與5mg/(kg·d) FK506、20mg/(kg·d) CsA的皮片存活時間相等，而腫瘤體積顯著減小(P<0.05)。與對照組相比，實驗劑量RPM顯著降低了荷瘤小鼠血清中的VEGF含量(P<0.05)，同時瘤組織內(nèi)的MVD顯著減少(P<0.05)。結論 體外實驗研究和動物實驗證實，RPM具有抗免疫排斥和抗腫瘤增殖的特點，可能在肝移植治療肝臟惡性腫瘤方面發(fā)揮優(yōu)勢。
【關鍵詞】 雷帕霉素;免疫抑制劑;肝癌;血管內(nèi)皮細胞生長因子
Inhibitory effect of rapamycin on proliferation of H22 hepatic cancer in mice
WU Zheng, L Yi, LIU Yuanxing, WANG Zuoren
Department of Hepatobiliary Surgery, the First Affiliated Hospital,Medical School of Xian Jiaotong University, Xian 710061, China醫(yī).學全.在.線網(wǎng)站gydjdsj.org.cn
ABSTRACT: Objective To explore rapamycin's inhibitory effect on proliferation of H22 hepatic cancer in mice. Methods In vitro study: H22 hepatic cancer cell lines were cultured with rapamycin, CsA, FK506, and 5FU, respectively, for 48h. The different drugs' inhibitory effect on proliferation was determined through MTT. The influences of different agents on the H22 hepatic cancer cell cycle were observed by flow cytometry. The vascular endothelial growth factor (VEGF) concentration of the supernatant fluid of the cultured H22 hepatic cancer cell was detected by ELISA. In vivo study: C57BL/6 to Balb/c mice allogenic skin transplant was established, and the H22 hepatic cancer cell was implanted under skin. Rapamycin, CsA, FK506 and 5FU were fed to the mice, respectively. The effect of different immunosuppressors on the survival of skin graft was observed while the proliferation of the transplant tumor was investigated. VEGF concentration of treated mice serum was examined by ELISA. The microvessel density of the transplanted tumor was observed through immunohistochemistry staining of CD34. Results The proliferation of the H22 hepatic cancer cells was inhibited by rapamycin at the concentration of 0.01-1mg/L. When the H22 hepatic cancer cells were cultured with different dose of rapamycin, the VEGF concentration of the supernatant fluid decreased significantly (P<0.05). The number of S phase cells decreased significantly compared to that of other agents (P<0.05). When the mice in different groups were fed with 1.5mg/(kg·d) and 4.5mg/(kg·d) rapamycin, the lengthened survival time of the skin grafts was similar to that in CsA and FK506 groups. But the tumor volume was smaller than that in CsA and FK506 groups (P<0.05). Compared to that in the control group, the VEGF concentration of mice serum decreased in rapamycin group (P<0.05), and the microvessel density of the transplant tumor was reduced greatly (P<0.05). Conclusion Rapamycin, as an immunosuppressor, significantly resists immunologic rejection and inhibits the proliferation of H22 hepatic cancer, thus having its advantage in treating malignant hepatic cancer with liver transplantation.
KEY WORDS: rapamycin; immunosuppressor; hepatic cancer; vascular endothelial growth factor (VEGF)
臨床資料表明，肝癌患者接受肝移植治療的總體療效還不能令人滿意，移植術后肝癌復發(fā)的問題一直是肝癌肝移植需要解決的關鍵問題之一。肝癌肝移植術后復發(fā)與圍手術期應用的免疫抑制劑有相當密切的聯(lián)系。本研究將探討免疫抑制劑雷帕霉素(rapamycin, RPM)對H22肝癌細胞生長增殖的影響，為防治肝移植術后肝癌復發(fā)的研究提供基礎理論依據(jù)。
