网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
miRNA-30c对鼻咽癌细胞放疗敏感性的影响
作者:金爱燕  李丽萍  朱虹  左文娜  王洪芹 
单位:沧州市中心医院 耳鼻喉科, 河北 沧州 061000
关键词:微小RNA-30c 鼻咽癌 放疗敏感性 
分类号:R739.6
出版年·卷·期(页码):2021·49·第八期(892-896)
摘要:

目的:探讨miR-30c对鼻咽癌放疗敏感性的影响。方法:鼻咽癌放疗耐受细胞(CNE1)细胞分为3组:空白对照组,阴性对照组,mimic组;放疗敏感细胞(CNE2)细胞分为3组:空白对照组,阴性对照组,antagomic组。荧光定量聚合酶链反应(PCR)检测miR-30c表达水平,克隆形成实验检测细胞增殖,流式细胞仪检测凋亡。Western blot检测蛋白表达。结果:CNE1细胞的空白对照组、阴性对照组和mimic组miR-30c的2-ΔΔCt值分别为0.165±0.037、0.162±0.035和0.463±0.064,CNE2细胞的空白对照组、阴性对照组和antagomic组的2-ΔΔCt值分别为0.323±0.045、0.326±0.047和0.204±0.038。CNE1细胞mimic组miR-30c的2-ΔΔCt值显著高于空白对照组和阴性对照组(P<0.01),CNE2细胞antagomic组miR-30c的2-ΔΔCt值显著低于空白对照组和阴性对照组(P<0.01)。CNE1和CNE2细胞空白对照和阴性对照组间差异无统计学意义(P>0.05)。CNE1细胞miR-30c的2-ΔΔCt值显著低于CNE2细胞(P<0.01)。CNE1细胞的空白对照组、阴性对照组和mimic组克隆形成率分别为(27.44±4.26)%、(27.25±4.21)%和(18.32±3.04)%,CNE2细胞的空白对照组、阴性对照组和antagomic组克隆形成率分别为(10.26±2.52)%、(10.22±2.47)%和(23.63±4.15)%。CNE1细胞mimic组克隆形成率显著低于空白对照组和阴性对照组(P<0.01),CNE2细胞antagomic组克隆形成率显著高于空白对照组和阴性对照组(P<0.01)。CNE1和CNE2细胞空白对照和阴性对照组间差异无统计学意义(P>0.05)。CNE1细胞的空白对照组、阴性对照组和mimic组细胞凋亡率分别为(8.16±1.31)%、(7.94±1.28)%和(28.46±4.17)%,CNE2细胞的空白对照组、阴性对照组和antagomic组细胞凋亡率分别为(33.16±5.42)%、(33.61±5.52)%和(20.14±3.26)%。CNE1细胞mimic组细胞凋亡率显著高于空白对照组和阴性对照组(P<0.01),CNE2细胞antagomic组细胞凋亡率显著低于空白对照组和阴性对照组(P<0.01)。CNE1和CNE2细胞空白对照组和阴性对照组间差异无统计学意义(P>0.05)。CNE1细胞的空白对照组、阴性对照组和mimic组LC3Ⅱ蛋白的相对灰度值分别为0.684±0.107、0.676±0.113和0.184±0.034,CNE2细胞的空白对照组、阴性对照组和antagomic组LC3 Ⅱ蛋白的相对灰度值分别为0.112±0.023、0.108±0.021和0.376±0.045。CNE1细胞mimic组LC3Ⅱ蛋白的相对灰度值显著低于空白对照组和阴性对照组(P<0.01),CNE2细胞antagomic组LC3Ⅱ蛋白的相对灰度值显著高于空白对照组和阴性对照组(P<0.01)。CNE1和CNE2细胞空白对照和阴性对照组间差异无统计学意义(P>0.05)。结论:miR-30c可以显著抑制鼻咽癌细胞放射线照射时自噬的激活,显著提高鼻咽癌细胞的放疗敏感性。

Objective: To explore the effects of miR-30c on radiosensitivity of nasopharyngeal carcinoma cells. Methods: CNE1 cells were divided into blank control, negative control andmimic group. CNE2 cells were divided into blank control, negative control andantagomic group. The expression of miR-30c was detected by real-time polymerase chain reaction(PCR) assay. The growth of cells was detected by clonogenic experiment. The apoptosis was detected by flow cytometry. The protein was detected by Western Blot analysis. Results: The 2-ΔΔCt values of miR-30c in blank control, negative control, mimic group of CNE1 were 0.165±0.037, 0.162±0.035, and 0.463±0.064, and that in blank control, negative control, antagomic group of CNE2 were 0.323±0.045, 0.326±0.047, and 0.204±0.038. The 2-ΔΔCt values of miR-30c in antagomic group were lower than that in blank control and negative control group(P<0.01). There was no significant difference between the blank control and the negative control group(P>0.05). The 2-ΔΔCt values of miR-30c in the mimic group was higher than that in the blank control and the negative control group(P<0.01). The clone formation rate in the blank control, the negative control, the mimic group of CNE1 were(27.44±4.26)%,(27.25±4.21)%, and(18.32±3.04)%, and it were(10.26±2.52)%,(10.22±2.47)%, and(23.63±4.15)% in the blank control, the negative control, the antagomic group of CNE2. The clone formation rate in mimic group was lower than that in the blank control and the negative control group(P<0.01). The clone formation rate in the antagomic group was higher than that in the blank control and the negative control group(P<0.01). There was no significant difference between the blank control and the negative control group(P>0.05). The apoptic rate in blank control, negative control, mimic group of CNE1 were(8.16±1.31)%,(7.94±1.28)%, and(28.46±4.17)%, and in blank control, negative control, antagomic group of CNE2 were(33.16±5.42)%,(33.61±5.52)%, and(20.14±3.26)%. The apoptic rate in mimic group was higher than that in the blank control and the negative control group(P<0.01). The apoptic rate in the antagomic group was lower than that in the blank control and the negative control group(P<0.01). There was no significant difference between the blank control and the negative control group(P>0.05). The relative gray value of LC3Ⅱ in the blank control, the negative control, the mimic group of CNE1 were 0.684±0.107, 0.676±0.113, and 0.184±0.034, andit were 0.112±0.023, 0.108±0.021, and 0.376±0.045 in blank control, negative control, antagomic group of CNE2. The relative gray value of LC3Ⅱ in mimic group was lower than that in the blank control and the negative control group(P<0.01). The relative gray value of LC3Ⅱ in antagomic group was higher than that in the blank control and the negative control group(P<0.01). There was no significant difference between the blank control and the negative control group(P>0.05).Conclusion: miR-30c can significantly inhibit the activation of autophagy in nasopharyngeal carcinoma cells during irradiation, and significantly improve the radiosensitivity of nasopharyngeal carcinoma cells.

参考文献:

[1] 梁锌, 杨剑, 高婷, 等.中国鼻咽癌流行概况[J]. 中国肿瘤, 2016, 25(11):835-840.
[2] 唐华燕, 吴辰, 李柔.肿瘤放射增敏剂在鼻咽癌放射治疗中的应用价值[J]. 中国当代医药, 2018, 25(23):109-111.
[3] 彭先兵, 戴润芝.Epstein Barr病毒编码的microRNA bart7调节鼻咽癌放疗敏感性的机制[J]. 山西医科大学学报, 2019, 50(6):834-838.
[4] XU X, KONG X, LIU T, et al. Metastasis-associated protein 1, modulated by miR-30c, promotes endometrial cancer progression through AKT/mTOR/4E-BP1 pathway[J]. Gynecol Oncol, 2019, 154(1):207-217.
[5] 乔国梁, 郭敛容, 方黎.抑制Polo样激酶1对鼻咽癌细胞CNE-1和CNE-2辐射敏感性的影响[J]. 中国实用医药, 2018, 13(11):194-195.
[6] WANG Z, MAO J W, LIU G Y, et al. MicroRNA-372 enhances radiosensitivity while inhibiting cell invasion and metastasis in nasopharyngeal carcinoma through activating the PBK-dependent p53 signaling pathway[J]. Cancer Med, 2019, 8(2):712-728.
[7] 张美婷, 山顺林, 耿炜.癌细胞放射抗拒性机制的研究进展[J]. 东南大学学报(医学版), 2017, 36(1):116-119.
[8] LI J, TU Z, SHEN Z, et al. Quantitative measurement of optical attenuation coefficients of cell lines CNE1, CNE2, and NP69 using optical coherence tomography[J]. Lasers Med Sci, 2013, 28(2):621-625.
[9] LI X H, HA C T, FU D, et al. Micro-RNA30c negatively regulates REDD1 expression in human hematopoietic and osteoblast cells after gamma-irradiation[J]. PLoS One, 2012, 7(11):e48700-48709.
[10] HE H, LIN K, SU Y, et al. Overexpression of β-Catenin Decreases the Radiosensitivity of Human Nasopharyngeal Carcinoma CNE-2 Cells[J]. Cell Physiol Biochem, 2018, 50(5):1929-1944.
[11] 陈瑶, 李思维.鼻咽癌放疗敏感性的研究进展[J]. 华夏医学, 2017, 30(1):167-171.
[12] LI B X, CAO X F, WENG C Y, et al. The regulatory effects of autophagy to the CNE2 cells radio-sensitization[J]. Cell Mol Biol, 2016, 62(13):12-14.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 510042 位访问者


 ©《现代医学》编辑部
联系电话:025-83272481;83272479
电子邮件: xdyx@pub.seu.edu.cn

苏ICP备09058541