| 陈志达,钟渊福,陈云萍,等.热疗联合特异性沉默趋化因子受体4对骨肉瘤增殖和侵袭的影响.骨科,2020,11(2): 143-148. |
| 热疗联合特异性沉默趋化因子受体4对骨肉瘤增殖和侵袭的影响 |
| Synergistic effect of hyperthermia combined with chemokine receptor 4 knockdown on proliferation and invasion of osteosarcoma |
| 投稿时间：2019-08-30 |
| DOI：10.3969/j.issn.1674-8573.2020.02.010 |
| 中文关键词: 骨肉瘤 CXCR4 细胞增殖 侵袭 |
| 英文关键词: Osteosarcoma CXCR4 Cell proliferation Invasion |
| 基金项目:国家自然科学基金（81402217）；漳州市自然科学基金（ZZ2018J10） |
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| 中文摘要: |
| 目的 观察热疗联合特异性siRNA沉默趋化因子受体4（chemokine receptor 4, CXCR4）对骨肉瘤增殖和侵袭的影响。方法 培养人骨肉瘤细胞株MG-63至对数生长期，接种于6孔板中，分为三组，空白组细胞不进行任何处理常规培养，control siRNA组以control siRNA转染细胞，CXCR4 siRNA组以CXCR4 siRNA转染细胞，采用Western blot检测各组CXCR4的表达变化，评估CXCR4 siRNA转染效能。细胞及体内实验设置control siRNA组（对照组）、热疗联合CXCR4 siRNA组（联合组）、CXCR4 siRNA组（沉默组）和热疗组，以CXCR4 siRNA转染沉默组和联合组的细胞，control siRNA转染对照组细胞，热疗组和联合组进行热疗处理。采用实时荧光定量聚合酶链反应（real-time quantitative polymerase chain reaction, RT-qPCR）和Western blot检测各组细胞CXCR4的表达变化。CCK-8法和Transwell侵袭实验检测转染后MG-63细胞的增殖能力和侵袭能力变化。流式细胞技术检测转染后MG-63细胞的周期变化。体外裸鼠成瘤实验和肿瘤生长曲线观察转染后MG-63细胞增殖和裸鼠骨肉瘤生长情况。结果 CXCR4 siRNA可以有效沉默MG-63细胞中CXCR4 蛋白的表达。热疗联合CXCR4 siRNA有效沉默MG-63细胞中CXCR4的表达并能抑制MG-63细胞增殖和侵袭，阻滞细胞周期于G0/G1期（P均＜0.05）。体内实验结果显示，从第14天起，联合组裸鼠骨肉瘤体积逐渐小于对照组、沉默组和热疗组，差异有统计学意义（P均＜0.05）。联合组的瘤重也低于对照组、沉默组和热疗组，差异有统计学意义（P均＜0.05）。结论 热疗联合特异性沉默CXCR4抑制骨肉瘤的增殖和侵袭，可作为一个有效的骨肉瘤基因治疗新方法。 |
| 英文摘要: |
| Objective To detect the synergistic effect of hyperthermia combined with chemokine receptor 4 (CXCR4) knockdown on the proliferation and invasion of osteosarcoma. Methods Osteosarcoma cells (human osteosarcoma cell line MG-63) were divided into three groups. The blank group cells were cultured without any treatment. The control siRNA group cells were transfected with control siRNA, and the CXCR4 siRNA group cells were transfected with CXCR4 siRNA. Western blotting was used to detect the expression of CXCR4 in each group and the transfection efficiency of CXCR4 siRNA was evaluated. The cell and in vivo experiments were set up with control siRNA group, hyperthermia combined with CXCR4 siRNA group, CXCR4 siRNA group and hyperthermia group. Cells in CXCR4 siRNA group and hyperthermia combined with CXCR4 siRNA group were transfected with CXCR4 siRNA. The control siRNA group was transfected with control siRNA. Hyperthermia group and hyperthermia combined with CXCR4 siRNA group were treated with hyperthermia. CXCR4 expression was detected by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting. CCK-8 method was used to detect the proliferation of MG-63 cells after transfection. Transwell invasion method was used to detect the invasion ability of MG-63 cells after transfection. Flow cytometry was used to detect the cell cycle effect of MG-63 cells after transfection. In vitro nude mice tumor formation experiment and tumor growth curve were used to detect the osteosarcoma cell proliferation and nude mouse osteosarcoma growth after transfection. Results CXCR4 siRNA could effectively silence the expression of CXCR4 in MG-63 cells. Hyperthermia combined with CXCR4 could effectively silence the expression of CXCR4 in osteosarcoma, inhibit the proliferation and invasion of osteosarcoma cells, and block the cell cycle at G0/G1 phase (all P＜0.05). The volume of osteosarcoma in nude mice treated with hyperthermia combined with CXCR4 siRNA was gradually decreased as compared with that in control siRNA group, CXCR4 siRNA group and hyperthermia group from the 14th day (all P＜0.05). The tumor weight of hyperthermia combined with CXCR4 siRNA group was also significantly reduced as compared with that in control siRNA group, CXCR4 siRNA group and hyperthermia group (P＜0.05). Conclusion The findings suggest that hyperthermia combined with CXCR4 knockdown can inhibit the proliferation and invasion of osteosarcoma, which may therefore serve as a new effective gene therapeutic target for osteosarcoma. |
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