普鲁卡因调节MAPK/ERK信号通路对甲状腺癌B-CPAP细胞增殖、迁移、侵袭及凋亡的影响实验研究OA
Impacts of procaine on proliferation,migration,invasion and apoptosis of thyroid cancer B-CPAP cells by regulating MAPK/ERK pathway
目的:探究普鲁卡因(PCA)调节丝裂原活化蛋白激酶(MAPK)/细胞外信号调节激酶(ERK)通路对甲状腺癌B-CPAP细胞增殖、迁移、侵袭及凋亡的影响.方法:培养人甲状腺癌B-CPAP细胞,分为对照组(CK组)、低剂量PCA组(L-PCA组)、中剂量PCA组(M-PCA组)、高剂量PCA组(H-PCA组)和H-PCA+MAPK激活剂茴香霉素(Anisomycin)组(H-PCA+Anisomycin组).克隆形成实验检测B-CPAP细胞增殖;Transwell实验检测B-CPAP细胞迁移、侵袭;吖啶橙(AO)/溴化乙啶(EB)染色检测B-CPAP细胞凋亡;蛋白质印记法(Western blot)检测增殖、迁移、侵袭、凋亡及MAPK/ERK通路相关蛋白表达;裸鼠移植瘤实验(对照组与PCA组,每组8只)检测移植瘤体积和重量;免疫组化染色检测细胞周期蛋白D1(Cyclin D1)、Ki-67抗原(Ki-67)、波形蛋白(Vimentin)、上皮型钙黏蛋白(E-cadherin)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax).结果:L、M、H-PCA组克隆形成数、迁移细胞数、侵袭细胞数,Cyclin D1、Ki-67、Vimentin、Bcl-2蛋白表达,以及p-P38 MAPK/P38 MAPK、p-ERK1/2/ERK1/2 比值较 CK 组降低,细胞凋亡率及 E-cadherin、Caspase-3、Bax 蛋白表达较CK组升高(均P＜0.05).H-PCA+Anisomycin组克隆形成数、迁移细胞数、侵袭细胞数,Cyclin D1、Ki-67、Vimen-tin、Bcl-2蛋白表达,以及p-P38 MAPK/P38 MAPK、p-ERK1/2/ERK1/2比值较H-PCA组升高,细胞凋亡率及E-cadherin、Caspase-3、Bax蛋白表达较H-PCA组降低(均P＜0.05).PCA组裸鼠移植瘤体积、重量及Cyclin D1、Ki-67、Vimentin、Bcl-2蛋白阳性细胞率较对照组降低,E-cadherin、Caspase-3、Bax蛋白阳性细胞率较对照组升高(均P＜0.05).结论:PCA对甲状腺癌B-CPAP细胞增殖、迁移、侵袭的抑制及凋亡的促进,可能是通过抑制MAPK/ERK通路实现的.
Objective:To explore the impacts of procaine(PCA)on the proliferation,migration,invasion and ap-optosis of thyroid cancer B-CPAP cells by regulating the mitogen-activated protein kinase(MAPK)/extracellular regulated protein kinase(ERK)pathway.Methods:Human thyroid cancer cell B-CPAP was cultured and assigned in-to control group(CK group),low-dose PCA group(L-PCA group),medium-dose PCA group(M-PCA group),high-dose PCA group(H-PCA group),and the H-PCA+MAPK activator anisomycin group(H-PCA+Anisomycin group).Cloning formation experiment was used to detect the proliferation of B-CPAP cells.Transwell assay was im-plemented to detect migration and invasion of B-CPAP cells.Acridine orange(AO)/ethidium bromide(EB)staining was implemented to detect apoptosis of B-CPAP cells.Western blot was implemented to detect proliferation,migra-tion,invasion,apoptosis,and MAPK/ERK pathway-related protein expression.Nude mouse transplantation tumor experiment(control group and PCA group,8 nude mice in each group)was used to detect the volume and weight of transplanted tumors.Immunohistochemical staining was implemented to detect the expression of Cyclin D1,Ki-67,Vimentin,epithelial cadherin(E-cadherin),Cysteine aspartic acid specific protease-3(Caspase-3),B-cell lymphoma-2(Bcl-2),and Bcl-2 associated X protein(Bax).Results:Compared with the CK group,the L-,M-,and H-PCA groups had a decrease in the number of clones formed,the number of migrating cells,the number of invading cells,and the expression of Cyclin D1,Ki-67,Vimentin,Bcl-2 proteins,and the ratios of p-P38 MAPK/P38 MAPK and p-ERK1/2/ERK1/2,and had an increase in apoptosis rate,and the expression of E-cadherin,Caspase-3,and Bax proteins(all P＜0.05).Compared with the H-PCA group,the H-PCA+anisomycin group had an increase in the number of clones formed,the number of migrating cells,the number of invading cells,and the expression of Cyclin D1,Ki-67,Vimen-tin,Bcl-2 proteins,and the ratios of p-P38 MAPK/P38 MAPK and p-ERK1/2/ERK1/2,and had a decrease in apop-tosis rate,the expression of E-cadherin,Caspase-3,and Bax proteins(all P＜0.05).Compared with the control group,the PCA group had a decrease in volume and weight of the transplanted tumors,and Cyclin D1,Ki-67,Vimen-tin,and Bcl-2 positive cell rates,and an increase in E-cadherin,Caspase-3,and Bax positive cell rates(all P＜0.05).Conclusion:The inhibitory effect of PCA on proliferation,migration and invasion as well as the pro-apoptotic effect on thyroid cancer B-CPAP cells may be achieved by inhibiting the MAPK/ERK pathway.
何概易南;苏元元;顾新华
苏州京东方医院药剂科,江苏苏州 215200苏州京东方医院药剂科,江苏苏州 215200苏州市立医院普外科,江苏苏州 215002
医药卫生
甲状腺癌普鲁卡因丝裂原活化蛋白激酶/细胞外信号调节激酶通路增殖迁移侵袭凋亡
Thyroid cancerProcaineMitogen-activated protein kinase/extracellular signal-regulated kinase pathwayProliferationMigrationInvasionApoptosis
《陕西医学杂志》 2026 (6)
748-753,760,7
江苏省基础研究计划(自然科学基金)专项资金资助项目(BK20230108)
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