首页|POSTN对人下咽癌细胞增殖、凋亡和EMT的影响

POSTN对人下咽癌细胞增殖、凋亡和EMT的影响

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目的 研究骨膜蛋白(Periostin,POSTN)对人下咽癌细胞增殖、凋亡和上皮-间质转化(epithelial-mesenchymal transi-tion,EMT)的影响和机制.方法 通过TIMER2数据库分析POSTN基因在头颈部肿瘤中的表达情况.培养人下咽癌FaDu细胞,构建转染si-POSTN的实验组(si-POSTN1组、si-POSTN2组、si-POSTN3组)和转染阴性对照序列si-NC的对照组(NC组),然后通过PCR实验筛选出POSTN基因敲低效率较高的两组,用于后续的实验.CCK-8和细胞集落形成实验检测增殖情况;划痕和Transwell实验检测迁移和侵袭能力;Western blot实验检测凋亡相关蛋白(Bax、Bcl-2)和EMT相关蛋白(N-cadherin、E-cadherin、Vimentin、Snail)的表达情况;PCR 实验检测 TGF-β/Smad 通路相关基因(TGF-β1、SMAD2、SMAD3)mRNA 的表达情况.结果 PCR实验发现si-POSTN1组和si-POSTN2组POSTN mRNA表达量比si-POSTN3组低(P<0.05),故选择si-POSTN1和si-POSTN2序列进行后续实验.与NC组相比,si-POSTN1组和si-POSTN2组POSTN的蛋白表达量下降(P<0.001).与NC组相比,si-POSTN1组和si-POSTN2组细胞增殖、迁移和侵袭能力均明显降低(P<0.05).与NC组相比,si-POSTN1组和si-POSTN2组凋亡相关蛋白Bax的表达量上升而Bcl-2的表达量减少(P<0.01),EMT相关蛋白E-cadherin的表达量增加而N-cadherin、Vimentin 和 Snail 的表达量均减少(P<0.05).与 NC 组相比,si-POSTN1 组和 si-POSTN2 组 TGF-β1、SMAD2、SMAD3的mRNA表达量均降低(P<0.01).结论 POSTN会增强下咽癌细胞迁移、侵袭、增殖的能力,促进EMT的进程,抑制细胞凋亡,其机制可能是通过TGF-β/Smad通路来调控.
Effects of POSTN on proliferation,apoptosis and EMT of human hypopharyngeal carcinoma cells
Objective To investigate the effect of Periostin(POSTN)on proliferation,apoptosis and epithelial-mesenchymal transition(EMT)in human hypopharyngeal carcinoma cells and its mechanisms.Methods The expression of POSTN in head and neck carci-noma was analyzed by TIMER2 database.Human hypopharyngeal carcinoma FaDu cells were cultured and transfected with different si-POSTN(si-POSTN1 group,si-POSTN2 group,si-POSTN3 group),and negative control sequence si-NC(NC group),respectively.Then two groups with higher POSTN knockdown efficiency were screened by PCR for the subsequent experiments.CCK-8 and colony formation assay were used to detect the cell proliferation.Cell scratch test and Transwell assay were used to detect the abilities of cell migration and invasion.Western blot was used to detect the expressions of apoptosis-related proteins(Bax,Bcl-2)and EMT-related proteins(N-cadherin,E-cadherin,Vimentin,Snail).PCR was used to detect the mRNA expressions of TGF-β/Smad pathway-related genes(TGF-β1,SMAD2,SMAD3).Results PCR results showed that the mRNA expression of POSTN in si-POSTN1 group and si-POSTN2 group was lower than that in si-POSTN3 group(P<0.05),so si-POSTN1 and si-POSTN2 were screened for the subsequent experiments.The protein expression of POSTN in si-POSTN1 group and si-POSTN2 group was lower than that in NC group(P<0.001).Compared with NC group,the cell proliferation,migration and invasion abilities were significantly reduced in si-POSTN1 group and si-POSTN2 group(P<0.05).Compared with NC group,the protein expression of Bax was increased while the protein expression of Bcl-2 was decreased in si-POSTN1 group and si-POSTN2 group(P<0.01),the expression of E-cadherin was increased while the expressions of N-cadherin,Vimentin and Snail were decreased in si-POSTN1 group and si-POSTN2 group(P<0.05).Compared with NC group,the expressions of TGF-β1,SMAD2 and SMAD3 were decreased in si-POSTN1 group and si-POSTN2 group(P<0.01).Conclusion POSTN can promote the migration,invasion and proliferation abilities of human hypopharyngeal carcinoma cells,promote the process of EMT,inhibit the cell apoptosis,which may be regulated by TGF-β/Smad pathway.

hypopharyngeal carcinomaPOSTNEMTproliferationapoptosis

马赛菲、邵渝欢、王文忠

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蚌埠医科大学第一附属医院耳鼻咽喉头颈外科,蚌埠 233000

下咽癌 POSTN 上皮-间质转化 增殖 凋亡

安徽省高等学校自然科学研究项目

KJ2020A0568

2024

山西医科大学学报
山西医科大学

山西医科大学学报

CSTPCD
影响因子:0.931
ISSN:1007-6611
年,卷(期):2024.55(3)
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