β-榄香烯对人肺腺癌裸鼠移植瘤放疗增敏作用与HIF-1α CAⅨ的相关性研究

  • 摘要: 目的:建立人肺腺癌A549细胞株裸鼠移植瘤模型, 研究β-榄香烯对移植瘤的放疗增敏效应, 寻找最佳增敏剂量, 并探讨放疗增敏机制是否与HIF-1α、 CAⅨ具有相关性。方法: 采用细胞悬液接种法建立裸鼠移植瘤模型, 待移植瘤体积达0.8~1.0 cm3时, 将裸鼠随机分组, 观测各组移植瘤的体积倍增时间。通过计算增敏系数, 获得最佳增敏剂量。RT-PCR及免疫组化方法检测HIF-1α表达, 免疫组化检测CAⅨ的表达。结果: 成功建立裸鼠移植瘤模型。25、 45、 100 mg/kg三种浓度β-榄香烯的增敏系数EF值分别为0.84、 1.24、 2.04, 最佳增敏剂量为45 mg/kg。药物组、 放疗组与空白组比较, HIF-1αmRNA表达明显增强 (P<0.01), 药物组与放疗组之间HIF-1αmRNA表达没有差异 (P>0.05), 联合组与其他各组比较, HIF-1αmRNA表达明显降低 (P<0.01)。各组HIF-1α的免疫组化情况: 药物组与空白组比较HIF-1α表达略增强 (P>0.05); 放疗组与空白组比较HIF-1α表达明显增强 (P<0.05), 联合组与其他各组比较HIF-1α表达明显降低 (P<0.01)。各组CAⅨ的免疫组化情况: 放疗组与空白组比较CAⅨ表达略增强 (P>0.05), 药物组与空白组比较CAⅨ表达增强 (P<0.05), 联合组与其他各组较, CAⅨ表达明显降低 (P<0.05)。结论: 本研究证实β-榄香烯的最佳增敏剂量为45 mg/kg, 与放疗联合明显抑制HIF-1α、 CAⅨ的表达, 提示β-榄香烯通过改善乏氧状况达到放射增敏的作用, 值得进一步研究。

     

    Abstract: Radiosensitization by β-elemene of Transplanted Lung Adenocarcinoma Cells in Nude Mice isRelated to the Expression of HIF-1α and CAⅨGuoquan LI1, Bingbing XIE1, Yinghai CHEN1, Xiaolong LI2, Lijuan ZOU1Correspondence to: Lijuan ZOU, E-mail: zoulijuan1963@sina.com1Department of Radiation Oncology, Second Affiliated Hospital of Dalian Medical University, Dalian 116027, China2Radiation Oncology Center, People's Liberation Army No. 323 Hospital, Xi'an 710054, ChinaThis work was supported by National Natural Science Foundation of China (No.3077064 6) and Innovative Team Project of LiaoningProvincial Education Department (No.2008T031)Abstract Objective: To establish a nude mouse model transplanted with the A549 lung adenocarcinoma cell line, to study thechange in radiosensitivity of transplanted tumors treated by β-elemene, to obtain the optimal dose to enhance the radiosensitivity oftransplanted tumors, and to investigate the association between radiosensitization by β-elemene and the expression of HIF-1α and CAⅨ. Methods: The nude mouse model transplanted with A549 lung adenocarcinoma cells was established by inoculating with a cell sus-pension. When the transplanted tumor size reached 0.8~1.0 cm3, the nude mice were randomly grouped ( 5 per group ) and the volumedoubling time was observed. The EF values were calculated and the optimal dose to enhance the radiosensitivity of transplanted tumorswas obtained. The expression of HIF-1α was assessed by RT-PCR and immunohistochemistry. The expression of CAⅨ was detected byimmunohistochemistry. Results: A nude mouse model transplanted with the A549 lung adenocarcinoma cell line was successfully estab-lished. The EF were 0.84 ( 25 mg/kg β-elemene ), 1.24 ( 45 mg/kg β-elemene ) and 2.04 ( 100 mg/kg β-elemene ) and the optimal doseto enhance radiosensitivity of the transplanted tumor was 45 mg/kg. According to RT-PCR the expression of HIF-1α was significantlyincreased in the groups treated with β-elemene or radiation alone compared with the control group ( P < 0.01 ), and there was no differ-ence between the β-elemene and radiation groups ( P > 0.05 ). According to RT-PCR the expression of HIF-1α was greatly decreased inthe co-treatment group compared with the control group ( P < 0.01 ). The results of immunohistochemistry of HIF-1α showed no differ-ence between the group treated with β-elemene alone and the control group ( P > 0.05 ), the expression of HIF-1α in the group treatedwith radiation alone was higher than that in the control group ( P < 0.05 ), and the co-treatment group had the lowest expression of allgroups ( P < 0.01 ). The immunohistochemistry results showed that the expression of CAⅨ in the group treated with radiation alone was slightly higher than that of the control group ( P > 0.05 ), while the expression in the group treated with β-elemene alone was signif-icantly higher than that of the control group ( P < 0.05 ). Immunohistochemistry also revealed that the expression of CAⅨ in theco-treatment group was the lowest of all of the groups ( P < 0.05 ). Conclusion: This study showed the optimal radiosensitization doseof β-elemene is 45 mg/kg. β-elemene combined with radiation apparently suppresses the expression of HIF-1α and CAⅨ, indicateingthat β-elemene may radiosensitize tumor cells by ameliorating hypoxic conditions in tumors. These results warrant further study.Keywords β-elemene; Transplanted tumor; Radiosensitization; HIF-1α; CAⅨ

     

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