长链非编码RNA肝癌高表达转录本对骨肉瘤细胞放射敏感性的影响  

Effects of LncRNA HULC on radiosensitivity of osteosarcoma cells

作　　者：丁帅[1] 高延征[1] 张广泉[1] 陈书连[1] 曹臣[1] 李白羽[2] Ding Shuai;Gao Yanzheng;Zhang Guangquan;Chen Shulian;Cao Chen;Li Baiyu(Department of Spine and Spinal Cord, Henan Province People′s Hospital, Zhengzhou 450003 China;Department of Cancer and Radiotherapy, Henan Province People′s Hospital, Zhengzhou 450003, China)

机构地区：[1]河南省人民医院脊柱脊髓外科,郑州450003 [2]河南省人民医院放疗科,郑州450003

出　　处：《中华放射医学与防护杂志》2019年第7期487-492,共6页Chinese Journal of Radiological Medicine and Protection

摘　　要：目的探讨长链非编码RNA(LncRNA)肝癌高表达转录本(HULC)对骨肉瘤细胞放射敏感性的影响。方法shRNA HULC慢病毒感染骨肉瘤细胞,以qRT-PCR测定干扰效果。以8 Gy X射线照射处理感染shRNA HULC慢病毒的骨肉瘤细胞,MTT、PI单染、Annexin V-FITC/PI双染法分别测定细胞增殖、周期和凋亡变化。Western blot检测细胞中p21、Cyclin D1、C-Caspase-3蛋白水平和胞浆、线粒体中细胞色素C(Cyt-C)蛋白水平。平板克隆实验检测骨肉瘤细胞的放射敏感性。结果shRNA HULC慢病毒感染可以明显下调骨肉瘤细胞中HULC表达水平。下调HULC或放射处理均可以抑制骨肉瘤细胞增殖[(100.00±9.65)% vs.(71.36±5.27)%、(63.48±5.93)%, t=4.512、5.585,P<0.05]、阻滞细胞周期[(50.15±5.14)% vs.(62.35±4.22)%、(66.05±5.23)%, t=3.177、3.756,P<0.05]和诱导细胞凋亡[(2.98±0.23)% vs.(22.61±3.26)%、(26.14±2.81)%,t=8.898、10.498,P<0.05],促进细胞中p21、C-Caspase-3蛋白表达并下调Cyclin D1蛋白水平,提高胞浆中Cyt-C蛋白水平并下调线粒体中Cyt-C蛋白水平。下调HULC联合放射对骨肉瘤细胞增殖[(71.36±5.27)%、(63.48±5.93)% vs.(49.32±5.76)%,t=4.890、2.967,P<0.05]、周期[(62.35±4.22)%、(66.05±5.23)% vs.(77.17±7.54)%,t=2.983、2.106,P<0.05]、凋亡[(22.61±3.26)%、(26.14±2.81)% vs.(36.21±3.26)%,t=6.164、4.564,P<0.05]及Cyclin D1、C-Caspase-3、Cyt-C蛋白表达影响作用更大。下调HULC后骨肉瘤细胞放射增敏比为1.432。结论下调HULC提高骨肉瘤细胞放射敏感性,这可能与协同放射阻滞细胞周期并诱导细胞凋亡有关。Objective To investigate the effect of LncRNA HULC on radiosensitivity of osteosarcoma cells. Methods Osteosarcoma cells OS732 was infected by shRNA HULC lentivirus, and the interference effect was determined by qRT-PCR. Osteosarcoma cells infected with shRNA HULC lentivirus were irradiated with 8 Gy X-rays. MTT, PI monochrome staining and Annexin V-FITC/PI double staining were used to detect cell proliferation, cell cycle and apoptosis, respectively. Western blot was used to detect the protein levels of p21, Cyclin D1, C-Caspase-3 and Cyt-C in cytoplasm and mitochondria. Plate cloning assay was used to evaluate cell radiosensitivity. Results The expression of HULC in osteosarcoma cells was significantly down-regulated by shRNA HULC lentivirus infection. Down-regulation of HULC or irradiation inhibited osteosarcoma cell proliferation [(100.00±9.65)% vs.(71.36±5.27)%,(63.48±5.93)%, t=4.512, 5.585, P<0.05], blocked cell cycle [(50.15±5.14)% vs.(62.35±4.22)%,(66.05±5.23)%, t=3.177, 3.756, P<0.05], induced cell apoptosis [(2.98±0.23)% vs.(22.61±3.26)%,(26.14±2.81)%, t=8.898, 10.498, P<0.05], promoted the expressions of p21 and Cyclin D1 in cells, down-regulated the level of C-Caspase-3 protein, increased the level of Cyt-C protein in cytoplasm, and down-regulated the level of Cyt-C protein in mitochondria. Downregulation of HULC combined with irradiation yield much more effects on cell proliferation inhibition [(71.36±5.27)%,(63.48±5.93)% vs.(49.32±5.76)%, t=4.890, 2.967, P<0.05], cell cycle arrest [(62.35±4.22)%,(66.05±5.23)% vs.(77.17±7.54)%, t=2.983, 2.106, P<0.05], apoptosis induction [(22.61±3.26)%,(26.14±2.81)% vs.(36.21±3.26)%, t=6.164, 4.564, P<0.05] and the expressions of p21, Cyclin D1, C-Caspase-3 and Cyt-C in osteosarcoma cells. The radiosensitization ratio of down-regulation of HULC was 1.432. Conclusions Down-regulation of HULC enhances radiosensitivity of osteosarcoma cells, which may be related to cell cycle arrest and apoptosis induction.

关 键 词：肝癌高表达转录本 放射 骨肉瘤 细胞凋亡 细胞周期 

分 类 号：R738[医药卫生—肿瘤] R730.55[医药卫生—临床医学]