[1]胡鹏刚,张昌明.枸杞皂苷通过调控Suv39H1/JAK2/STAT3通路对鼻咽癌细胞增殖、侵袭和凋亡的影响[J].现代检验医学杂志,2021,36(06):27-33.[doi:10.3969/j.issn.1671-7414.2021.06.006]
　HU Peng-gang,ZHANG Chang-ming.Lycium Barbarum Saponins Affect Proliferation, Invasion and Apoptosis of Nasopharyngeal Carcinoma Cells by Regulating the Suv39H1/JAK2/STAT3 Pathway[J].Journal of Modern Laboratory Medicine,2021,36(06):27-33.[doi:10.3969/j.issn.1671-7414.2021.06.006]
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枸杞皂苷通过调控Suv39H1/JAK2/STAT3通路对鼻咽癌细胞增殖、侵袭和凋亡的影响()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第36卷
期数:
2021年06期
页码:
27-33
栏目:
论　著
出版日期:
2021-12-15

文章信息/Info

Title:
Lycium Barbarum Saponins Affect Proliferation, Invasion and Apoptosis of Nasopharyngeal Carcinoma Cells by Regulating the Suv39H1/JAK2/STAT3 Pathway
文章编号:
1671-7414（2021）06-027-08
作者:
胡鹏刚; 张昌明
（空军军医大学第一附属医院西京医院耳鼻咽喉头颈外科，西安 710032)
Author(s):
HU Peng-gang;  ZHANG Chang-ming
（Otorhinolaryngology Head and Neck Surgery，Xijing Hospital, the First Affiliated Hospital of Air Force Military Medical University ，Xi’an 710032, China）
关键词:
枸杞皂苷; 鼻咽癌; Suv39H1; JAK2/STAT3信号通路
分类号:
R739.63；R730.43
DOI:
10.3969/j.issn.1671-7414.2021.06.006
文献标志码:
A
摘要:
目的　探究枸杞皂苷介导Suv39H1/JAK2/STAT3通路对鼻咽癌CNE-2细胞增殖、侵袭和凋亡的影响。方法　将人鼻咽癌CNE-2细胞株分为枸杞皂苷干预组（5，10，50 μmol/L枸杞皂苷）及对照组，采用CCK-8检测不同浓度枸杞皂苷作用0，24，48和72 h时CNE-2细胞增殖率，Annexin V-FITC/PI双染法流式细胞术检测枸杞皂苷干预48 h后CNE-2细胞凋亡率，Transwell小室实验检测CNE-2细胞侵袭能力，实时荧光定量聚合酶链式反应（qRT-PCR）与蛋白免疫印迹法（Western blot）分别检测组蛋白甲基化酶Suv39H1的表达水平；采用qRT-PCR和Western blot分别检测Suv39H1在鼻咽癌组织和癌旁组织，以及鼻咽癌细胞系（HONE1，C666-1，CNE-1和CNE-2）和人鼻咽癌上皮细胞（NP69）中的表达差异；构建Suv39H1过表达载体（pcDNA-Suv39H1）以及针对Suv39H1的小干扰RNA（Suv39H1 siRNA），分别转染于CNE-2细胞，检测细胞增殖、侵袭和凋亡；将pcDNA-Suv39H1载体转染于50 μmol/L枸杞皂苷干预的细胞中，分别检测细胞增殖、侵袭和凋亡。Western blot检测JAK2/STAT3通路相关蛋白p-JAK2和p-STAT3的表达。结果　与对照组相比，枸杞皂苷干预鼻咽癌CNE-2细胞呈剂量依赖性降低细胞增殖（F=12.030，P=0.002 5）和侵袭（F=4.807，P=0.031 5），增加细胞凋亡率（F=5.232，P=0.026 1），抑制Suv39H1蛋白表达（F=14.64，P=0.001 3）；与正常组织和NP69细胞相比，鼻咽癌组织和HONE1，C666-1，CNE-1，CNE-2细胞中Suv39H1表达水平显著升高（P<0.01）；转染pcDNA-Suv39H1组细胞增殖、侵袭率增强、凋亡率降低；转染Suv39H1 siRNA组细胞增殖、侵袭率降低，凋亡率升高，差异与对照组相比具有统计学意义（P<0.01） ；与50 μmol/L枸杞皂苷干预组相比，转染pcDNA-Suv39H1组细胞增殖、侵袭率增加、凋亡率降低，p-JAK2和p-STAT3蛋白表达升高（P<0.01）。结论　枸杞皂苷通过下调Suv39H1/JAK2/STAT3通路抑制鼻咽癌CNE-2细胞增殖、侵袭，促进凋亡。
Abstract:
Objective　To explore the effect of lycium barbarum saponins mediated Suv39H1/JAK2/STAT3 pathway on proliferation, invasion and apoptosis of nasopharyngeal carcinoma (NPC) CNE-2 cells. Methods　Human nasopharyngeal carcinoma CNE-2 cell lines were divided into lycium barbarum saponin treated groups (5, 10, 50μmol/L) and control group, and the proliferation rate of CNE-2 cells treated with different concentrations of lycium barbarum saponins at 0, 24, 48, 72 h was assessed by CCK-8 assay, followed by flow cytometry with annexin V-FITC/PI staining for 48 h after the apoptosis rate of CNE-2 cells, the invasion ability of CNE-2 cells was detected by Transwell chamber assay, and the expression levels of histone methylase Suv39H1 were determined by real-time PCR and Western blot, respectively. Differential expression of Suv39H1 was assessed in NPC tissues and adjacent noncancerous tissues, as well as in NPC cell lines (hone1, c666-1, CNE-1, and CNE-2) and human NPC epithelial cells (NP69), respectively. Suv39h1 overexpression constructs (pcDNA-Suv39h1) as well as small interfering RNA (Suv39h1 siRNA) against suv39h1 were transfected into CNE-2 cells and assessed for proliferation, invasion and apoptosis. pcDNA-Suv39h1 vector was transfected into cells treated with 50 μmol/L of lycium barbarum saponins to determine the proliferation, invasion and apoptosis abilities, respectively. The expression of JAK2/STAT3 pathway related proteins p-JAK2 and p-STAT3 was detected by Western blot assay. Results　Compared with the control group, lycium barbarum intervention in NPC CNE-2 cells decreased cell proliferation（F=12.030，P＜0.002 5）, invasion （F=4.807，P=0.031 5） and Suv39h1 protein level, and increased the apoptosis ratio in dose dependent manner（F=5.232，P=0.026 1）. Suv39h1 expression level was significantly higher in NPC tissues and HONE1, C666-1, CNE-1, and CNE-2 cells compared with normal tissues and NP69 cells (P < 0.01). CNE-2 cells transfected with pcDNA-Suv39h1 showed enhanced proliferation and invasion, and decreased apoptotic rate, while cells transfected with Suv39h1 siRNA displayed decreased proliferation and invasion and increased apoptotic rate, compared with control group (P< 0.01). Compared with the 50 μmol/L lycium barbarum saponins intervention group, the pcDNA-Suv39h1-transfected group showed increased cell proliferation and invasion rates, decreased apoptotic rates and increased protein expression of p-JAK2 and p-STAT3 (P < 0.01). Conclusion　Lycium barbarum saponins inhibit NPC CNE-2 cell proliferation, invasion, and promote apoptosis by downregulating the suv39h1/JAK2/STAT3 pathway.

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备注/Memo

备注/Memo:
作者简介：胡鹏刚（1980-），男，本科，主治医师，主要研究方向：分泌性中耳炎，E-mail:hupenggang123@126.com。通讯作者：张昌明（1970-），男，硕士，主治医师，主要研究方向：分泌性中耳炎。
更新日期/Last Update: 1900-01-01