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中国癌症防治杂志

中国癌症防治杂志 ›› 2023, Vol. 15 ›› Issue (5): 525-532.doi: 10.3969/j.issn.1674-5671.2023.05.09

• 基础研究 • 上一篇    下一篇

LGR4对鼻咽癌细胞增殖、凋亡、侵袭、迁移及上皮间质转化的影响

  

  1. 广西医科大学附属肿瘤医院放疗科；区域性高发肿瘤早期防治研究教育部重点实验室（广西医科大学）；广西鼻咽癌临床医学研究中心
  • 出版日期:2023-10-25 发布日期:2023-11-03
  • 通讯作者: 曲颂 E-mail：qusong2022@126.com
  • 基金资助:
    广西科技计划项目（桂科AD20297078）；广西医疗卫生适宜技术开发与推广应用项目（S2021020）；区域性高发肿瘤早期防治研究教育部重点实验室（GKE?ZZ202224）；区域性高发肿瘤早期防治研究教育部/广西重点实验室（GKE?ZZ202128）

Effects of LGR4 on proliferation,apoptosis,invasion,migration and epithelial mesenchymal transformation of nasopharyngeal carcinoma cells

  • Online:2023-10-25 Published:2023-11-03

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摘要： 目的 探讨富含亮氨酸重复G蛋白偶联受体⁃4（leucine⁃rich repeats containing G protein⁃coupled receptor 4，LGR4）对鼻咽癌C666⁃1细胞的增殖、凋亡、侵袭、迁移和上皮间质转化（epithelial⁃mesenchymal transition，EMT）的影响。 方法 利用韦恩图将GEO数据库测序数据GSE15903、GSE89804和GSE103611三组数据取交集得出差异基因。通过生物信息学分析LGR4在鼻咽癌组织及正常鼻咽上皮组织、鼻咽癌治疗后的转移或未转移组织中的表达，RT⁃qPCR和 Western blot检测LGR4在鼻咽癌C666⁃1细胞和正常鼻咽上皮细胞NP69中的表达水平。将鼻咽癌C666⁃1细胞进行慢病毒感染分为NC组（转染阴性对照）、KD1组和KD2组（敲低组），采用Western blot测定LGR4和上皮表型E⁃cadherin的表达，采用CCK⁃8、克隆形成实验、流式细胞术、划痕实验及Transwell 实验检测细胞增殖、凋亡、侵袭和迁移。结果 韦恩图显示GSE89804、GSE15903和GSE103611数据集的差异基因是LGR4。GSE12452数据分析结果显示，与正常鼻咽上皮组织相比，LGR4在鼻咽癌组织中表达上调（P<0.05）。GSE103611数据分析结果显示，与鼻咽癌治疗后未转移组织相比，LGR4在鼻咽癌治疗后转移组织中表达上调（P<0.05）。与NP69细胞相比，LGR4在鼻咽癌C666⁃1细胞中的mRNA和蛋白表达水平均显著上调（均P<0.001）。 与NC组相比，敲低LGR4可抑制鼻咽癌C666⁃1细胞增殖（均P<0.05）、侵袭及迁移能力（均P<0.001），促进凋亡和C666⁃1细胞中E⁃cadherin的表达（均P<0.001）。结论  LGR4在鼻咽癌C666⁃1细胞中表达上调，敲低LGR4抑制C666⁃1细胞的增殖、侵袭迁移，促进凋亡和上皮间质转化。

关键词: 鼻咽癌, 富含亮氨酸重复G蛋白偶联受体-4（LGR4）, 增殖, 凋亡, 侵袭, 迁移, 上皮间质转化

Abstract: Objective To investigate the effects of Leucine⁃rich repeats containing G protein⁃coupled receptor 4 (LGR4) on the proliferation, apoptosis, invasion, migration and epithelial mesenchymal transition (EMT) of nasopharyngeal carcinoma (NPC) cell lines C666⁃1. Methods The differential genes were obtained by intersection of GEO database sequencing data GSE89804, GSE15903 and GSE103611 using Venn diagram. The expression of LGR4 in NPC tissues, corresponding normal tissues，NPC tumor tissues with and without distant metastasis were analyzed by the bioinformatic analysis. The expression levels of LGR4 in NPC cell lines C666⁃1 and normal nasopharyngeal cell lines NP69 were detected by RT⁃qPCR and Western blot. NPC cell lines C666⁃1 were divided into the NC group ( negative control lentivirus), the KD1 group and the KD2 group (Knock Down). The expression of LGR4 and epithelial marker E⁃cadherin were detected by Western blot. Cell proliferation, apoptosis, invasion and migration were detected by CCK⁃8, clone formation assays, flow cytometry, scratch assay and Transwell assay. Results Venn diagram showed that the differential gene of GSE89804, GSE15903 and GSE103611 data sets was LGR4. The results of GSE12452 dataset showed that the LGR4 was up⁃regulated in NPC tissues by constrast with corresponding normal tissues (P<0.05). The results of GSE103611 dataset showed that the LGR4 was up⁃regulated in NPC tumor tissues with distant metastasis by contrast with NPC tumor tissues without distant metastasis (P<0.05). Compared with the mRNA and protein expression levels of NP69 cell lines, those of LGR4 were significantly up⁃regulated in NPC cell lines C666⁃1 (all P<0.001). Compared with the NC group, the knockdown of LGR4 inhibited the proliferation (all P<0.05), invasion and migration (all P<0.001), and promoted the apoptosis and protein expression of E⁃cadherin in C666⁃1 cell lines (all P<0.001). Conclusions  The expression of LGR4 is up⁃regulated in NPC cell lines C666⁃1. The knockdown of LGR4 could inhibit the proliferation, invasion, migration and promote apoptosis and EMT of NPC cell lines C666⁃1.

Key words: Nasopharyngeal carcinoma, Leucine-rich repeats containing G protein-coupledreceptor4(LGR4), Proliferation, Apoptosis, Invasion, Migration, Epithelial mesenchymal transition

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