长链非编码RNA CCAT1对人宫颈癌 XB1702细胞裸鼠移植瘤放射敏感性的影响

摘要

Objective To investigate the effect of long non-coding RNA (lnRNA) CCAT1 on the tumor sensitivity to radiotherapy in nude mice transplanted with human cervical cancer cells XB1702. Methods The human cervical cancer XB1702 cells were divided into 3 groups: control group, CCAT1 siRNA group and pcDNA3.1-CCAT1 group. The CCAT1 siRNA and pcDNA3.1-CCAT1 were transfected into the human cervical cancer cells XB1702 by the electroporation method and stable transfected cell line was selected and identified by the G418 method. After 48 h of transplantation, the expression levels of CCAT1 lnRNA were measured by RT-PCR. Cellular proliferation was tested by CCK8 method, and apoptosis were determined by the TUNEL technology. The development of human cervical tumors in the nude mice xenograft model was studied by up-regulating and down-regulating the CCAT1 expression in the cells combined with the X-ray irradiation. Results Interference of CCAT1 lnRNA expression in XB1702 cells significantly inhibited the cellular proliferation, increased apoptosis rate, and reduced the average xenografted tumor volume of the nude mice compared with control group. Up-regulation of CCAT1 lnRNA expression significantly increased the cellular proliferation and decreased apoptosis. The average xenograft tumor volume in this group had no obvious difference with the ones before the irradiation. Conclusion Down-regulating CCAT1 mRNA expression levels could inhibit the development of tumor and improve the tumor sensitivity to radiotherapy in nude mice transplanted with human cervical cancer cells XB1702, while up-regulating CCAT1 lnRNA expression levels could enhance the radiation resistance of tumor.%目的：探讨长链非编码 RNA（lncRNA）结肠癌相关转录因子1（CCAT1）对宫颈癌 XB1702细胞裸鼠移植瘤放射敏感性的影响。方法通过电穿孔法分别将 CCAT1 siRNA和重组表达载体 pcDNA3.1-CCAT1转染人宫颈癌 XB1702细胞后， G418筛选得到稳定转染细胞系，实验分为3组：CCAT1 siRNA 转染组、pcDNA3.1-CCAT1转染组和空白对照组。转染48 h后，RT-PCR检测细胞中 CCAT1 lncRNA表达水平；CCK8检测细胞增殖，TUNEL 法检测细胞凋亡影响；裸鼠移植瘤实验检测上调和下调 RNA CCAT1表达联合 X射线照射对宫颈癌的生长抑制作用。结果干扰 XB1702细胞中 CCAT1 lncRNA表达后， XB1702细胞增殖被抑制，细胞凋亡率增加；裸鼠移植瘤平均体积明显小于对照组（正常 XB1702细胞种植组）差异有统计学意义（P <0.05）；上调 XB1702细胞中 CCAT1 lncRNA表达后，XB1702细胞增殖能力明显增强，细胞凋亡率降低，差异有统计学意义（P <0.05），而上调组裸鼠皮下种植瘤平均体积较照射前无明显变化。结论下调 CCAT1 mRNA表达能抑制人宫颈癌 XB1702细胞裸鼠移植瘤的生长，增强瘤体的放射敏感性。