中华皮肤科杂志 ›› 2021, Vol. 54 ›› Issue (3): 220-225.doi: 10.35541/cjd.20200743

• 论著 • 上一篇    下一篇

RPL34基因敲低对人皮肤鳞状细胞癌SCL-1细胞的影响

张雪丽    郭燕    苏敏静    刘玉    黄艳平    李欣    孙志强    韩建文   

  1. 内蒙古医科大学附属医院皮肤性病科，呼和浩特  010010
  • 收稿日期:2020-07-24 修回日期:2021-01-06 发布日期:2021-03-02
  • 通讯作者: 韩建文 E-mail:hanjianwen1981@hotmail.com
  • 基金资助:
    国家自然科学基金（81660513）；内蒙古自治区自然科学基金（2018MS08030）；内蒙古自治区科技计划项目（201503004、2019GG082）；内蒙古自治区“草原英才”工程青年创新创业人才培养计划；中国科学院“西部之光”人才培养引进计划

Effect of ribosomal protein L34 gene knockdown on a human cutaneous squamous cell carcinoma cell line SCL-1

Zhang Xueli, Guo Yan, Su Minjing, Liu Yu, Huang Yanping, Li Xin, Sun Zhiqiang, Han Jianwen   

  1. Department of Dermatology and Venereology, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010010, China
  • Received:2020-07-24 Revised:2021-01-06 Published:2021-03-02
  • Contact: Han Jianwen E-mail:hanjianwen1981@hotmail.com
  • Supported by:
    National Natural Science Foundation of China（81660513）; Natural Science Foundation of Inner Mongolia Autonomous Region（2018MS08030）; Science and Technology Planning Project of Inner Mongolia Autonomous Region （201503004, 2019GG082）; The Young Innovative Talents Training Project of "Prairie Talents" of Inner Mongolia Autonomous Region; CAS "Light of West China" Program

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摘要： 【摘要】目的 研究ribosomal protein L34 (RPL34)基因敲低后对皮肤SCC细胞增殖、凋亡的影响。方法 首先通过免疫组化对14例皮肤cSCC患者和16例正常对照的组织进行RPL34表达分析。然后利用慢病毒感染皮肤鳞癌SCL-1 细胞RPL34表达进行基因敲低后，流式细胞仪检测细胞周期分布及凋亡，MTT检测细胞增殖，qPCR检测目的基因敲减效率，Western blot分析RPL34表达水平。计数资料用卡方检验进行比较，计量资料用均数±标准差表示，t检验比较2组间差异。α设定为0.05，P<0.05认为有统计学差异。结果免疫组化实验中，cSCC肿瘤组织中RPL34在细胞浆的表达明显高于正常对照皮肤组织（cSCC组细胞核表达评分为2.929±1.542，细胞浆表达评分为2.143±1.956，正常对照皮肤组织中细胞核表达评分为2.563 ±1.153，细胞浆表达评分为0.500±0.516。）。流式细胞仪分析检测细胞凋亡情况，在空白对照组凋亡率为4.58 %，而RPL34-shRNA组的凋亡率为9.42%（P<0.05）。细胞周期分布情况实验组（shRPL34）处于S期的细胞增多(P<0.05)，处于G1期的细胞减少(P<0.05)，处于G2/M期的细胞无显著变化(P＞0.05)。MTT检测细胞增殖，shRPL34组的SCL-1细胞OD490吸光率及吸收率变化倍数明显低于空白对照组(P<0.05)，提示具有活力的细胞数量明显低于对照组。qPCR结果中，经shRNA慢病毒感染后，实验组SCL-1细胞中RPL34基因在mRNA水平的表达量受到抑制，敲减效率达到85.1%（p<0.05）。结论 RPL34在cSCC中可以调节细胞增殖、凋亡，干扰细胞周期。

关键词: 癌, 鳞状细胞, 核糖体蛋白质类, RNA, 小分子干扰, 细胞周期, 细胞凋亡, RPL34, SCL-1细胞

Abstract: 【Abstract】Objective To investigate the effects of knockdown of the RPL34 gene on the proliferation and apoptosis of human cSCC cells. Methods First, RPL34 expression in the skin tissues of 14 patients with cSCC and 16 normal control subjects was analyzed using immunohistochemistry. Next, siRNA-lentivirus was used to knockdown RPL34 expression in the cSCC cells SCL-1. PI-FACS was used to measure cell cycle distribution and apoptosis, and MTT assay was also performed,the knockdown efficiency of target gene was detected by qPCR, and the expression level of RPL 34 was analyzed by Western blot. Chi-square test was used to compare the counting data, and the mean ± standard deviation was used to express the measurement data. When α was set to 0.05, P < 0.05 was considered to be statistically different. Results In immunohistochemistry, the expression of RPL 34 in cSCC tumor tissue was significantly higher than that in normal skin tissue (the score of nuclea expression in cSCC group was 2.929±1.542，the score of cytoplasm expression in cSCC group was 2.143±1.956, that in normal skin tissue was 2.563 ± 1.153, and that in cytoplasm was 0.500 ± 0.516, P < 0.05). The apoptosis rate was 4.58% in blank control group and 9.42% in RPL 34 shRNA group (P < 0.05). Cell cycle distribution in the experimental group (shRPL 34) increased in S phase (P < 0.05), decreased in G1 phase (P < 0.05), and no significant change was found in G2 / M phase (P > 0.05). MTT assay showed that the absorbance and absorbance of SCL-1 cells CD490 in shRPL 34 group were significantly lower than those in blank control group (P < 0.05), indicating that the number of viable cells in shRPL34group was significantly lower than that in control group. In qPCR results, the expression of RPL 34 gene in SCL-1 cells was inhibited by shRNA lentivirus infection, and the knockdown efficiency was 85.1% (P < 0.05). Conclusion RPL34 can regulate cell proliferation and apoptosis, as well as interfere with the cell cycle, in cSCC.

Key words: Carcinoma, squamous cell, Ribosomal proteins, RNA, small interfering, Cell cycle, Apoptosis, RPL34, SCL-1 cells