| 吴 婧,尹 丽,陈 猛.HAP1基因对人乳腺癌MDA-MB-231细胞裸鼠移植瘤放射敏感性的影响[J].中国肿瘤,2015,24(6):510-516. |
| HAP1基因对人乳腺癌MDA-MB-231细胞裸鼠移植瘤放射敏感性的影响 |
| The Effect of HAP1 on Radiosensitivity of Human Breast Carcinoma Cell MDA-MB-231 Xenograft in Nude Mice |
| 投稿时间：2014-11-19 |
| DOI：10.11735/j.issn.1004-0242.2015.06.A016 |
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| 中文关键词: 乳腺癌 放射敏感性 HAP1 细胞凋亡 |
| 英文关键词:breast Carcinoma Rdiosensitivity HAP1 Apoptosis |
| 基金项目: |
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| 中文摘要: |
| 摘 要：［目的］ 研究HAP1基因对人乳腺癌MDA-MB-231细胞裸鼠移植瘤放射敏感性的影响及作用机制。［方法］取对数生长期已构建好的MDA-MB-231/HAP1和MDA-MB-231/Pb稳转细胞株（5×106），接种于5周龄BALB/C 雄性裸鼠左腹股沟部皮下，建立人乳腺癌裸鼠移植瘤模型，21d裸鼠皮下肿瘤长径到达0.6~0.9cm时，分为4组（n=5）：MDA-MB-231/Pb组，MDA-MB-231/HAP1组，MDA-MB-231/Pb+放射组和MDA-MB-231/HAP1+放射组。放射组在第21d给予5Gy X线照射一次后继续饲养3周左右。期间每隔4d测量1次肿瘤的长径及短径，计算肿瘤体积。第6周时处死各组裸鼠统计瘤重。免疫组织化学方法分析Cleaved-caspase-3和Cleaved-caspase-9表达，TUNEL分析细胞凋亡。［结果］ MDA-MB-231/HAP1+放射组裸鼠肿瘤生长明显迟缓，平均体积小于其他3组（P均<0.01），MDA-MB-231/HAP1+放射组平均瘤重明显小于其他3组（P均<0.01）。TUNEL凋亡实验结果表明，MDA-MB-231/Pb组细胞平均凋亡率为3.27%±1.51%，MDA-MB-231/HAP1组为36.77%±3.78%，MDA-MB-231/Pb+放射组为46.27%±2.98%，MDA-MB-231/HAP1+放射组为83.07%±0.21%，MDA-MB-231/HAP1+放射组凋亡率明显高于其他3组（P<0.05）。免疫组织化学染色评分结果显示，MDA-MB-231/Pb组，MDA-MB-231/HAP1组，MDA-MB-231/Pb+放射组，MDA-MB-231/HAP1+放射组中凋亡蛋白Cleaved-caspase-3平均积分分别为1.67±0.47，6.00±1.63，7.00±1.41和14.67±1.89，凋亡蛋白Cleaved-caspase-9平均积分分别为2.00±0.82，4.67±0.94，5.33±0.94和11.00±1.41；Cleaved-caspase 3和cleaved-caspase 9在MDA-MB-231/HAP11+放射组中的水平明显高于其他3组（P<0.05）。［结论］ HAP1基因可能通过调控肿瘤细胞的凋亡来增强人乳腺癌细胞对放射治疗的敏感性。 |
| 英文摘要: |
| Abstract：［Purpose］ To study the effect and mechanism of HAP1 on radiosensitivity of human breast carcinoma cell MDA-MB-231 xenograft in nude mice.［Methods］ Constructed MDA-MB-231/HAP1 and MDA-MB-231/Pb stably transfected cells (5×106) were subcutaneously injected into nude mice to establish implanted tumor model of human breast cancer in nude mice. The nude mice were divided into 4 groups(5 mice with 20 days old per group) for MDA-MB-231/Pb group，MDA-MB-231/HAP1group，MDA-MB-231/Pb+ radiation group and MDA-MB-231/HAP1+ radiation group. The dose of IR was 5-Gy X-ray(once) and feeding for about 3 weeks. The long diameter and short diameter of the tumor were measured every 4 days. The tumor volume was weighted after 6 weeks. The Cleaved-caspase-3 and Cleaved-caspase-9 expression was analyzed by Immunohistochemical method. Apoptosis was analyzed by TUNEL assay.［Results］ The tumor volume of MDA-MB-231/HAP1 + radiation group was smallest than that in other three groups(all P<0.01). The weight of tumor in MDA-MB-231/HAP1 + radiation group was lowest than that in other three groups(all P<0.01). TUNEL showed that apoptosis rate of MDA-MB-231/Pb group，MDA-MB-231/HAP1 group，MDA-MB-231/Pb+ radiation group and MDA-MB-231/HAP1+ radiation group was 3.27%±1.51%，36.77%±3.78%，46.27%±2.98% and 83.07%±0.21% respectively，the apoptosis rate in MDA-MB-231/HAP1+ radiation group was significantly higher than that in other three groups (P<0.05). Immunohistochemistry showed that the expression of Cleaved-caspase-3 in MDA-MB-231/Pb group，MDA-MB-231/HAP1 group，MDA-MB-231/Pb+ radiation group and MDA-MB-231/HAP1+ radiation group was 1.67±0.47，6.00±1.63，7.00±1.41 and 14.67±1.89，respectively；and the expression of Cleaved-caspase-9 was 2.00±0.82，4.67±0.94，5.33±0.94 and 11.00 ±1.41，respectively. The expression of Cleaved-caspase-3 and Cleaved-caspase-9 in MDA-MB-231/HAP1+ radiation group was significantly higher than those in other three groups(P<0.05).［Conclusion］ HAP1 gene might enhance the sensitivity of human breast cancer cells to radiotherapy by regulating apoptosis. |
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