miRNA-30c对鼻咽癌细胞放疗敏感性的影响-现代医学

miRNA-30c对鼻咽癌细胞放疗敏感性的影响

单位：沧州市中心医院耳鼻喉科, 河北沧州 061000

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.

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