内质网PERK/eIF-2α信号通路在宫颈癌中的表达及其临床意义

Expression and Clinical Significance of the Endoplasmic Reticulum PERK/eIF-2α Signaling Pathway in Cervical Cancer

摘要

目的 探讨内质网应激蛋白激酶R样内质网激酶（stress protein kinase R-like endoplasmic reticulum kinase，PERK）/真核启动因子2α（eukaryotic initiation factor 2α，eIF-2α）信号通路在宫颈上皮内瘤变（cervical intraepithelial neoplasia，CIN）和宫颈癌（cervical carcinoma，CC）中的表达及其对肿瘤进程的影响。方法 收集2022年1月至2023年4月在南京明基医院进行诊治的CC患者的宫颈活检标本58例为宫颈癌组，选取同期52例宫颈上皮内瘤变（CIN Ⅱ级、CIN Ⅲ级）作为对照组，选取同期34例健康子宫患者的宫颈组织为正常组。Western blot法检测内质网介导相关蛋白的表达；免疫组织化学法检测PERK/eIF-2α在CC中的染色状况。将HeLa细胞分为对照组、pcDNA3.1-NC组、过表达PERK组、sh-NC组、干扰PERK组，采用CCK-8法检测细胞增殖活性，免疫荧光染色观察细胞内上皮型钙黏蛋白（E-cadherin）表达，应用流式细胞术检测细胞凋亡与周期分布，Transwell实验评估细胞迁移与侵袭能力。结果 随着CIN分级程度的增加，PERK/eIF-2α表达升高（P < 0.05），且CC组织PERK/eIF-2α表达显著高于CIN组织（P < 0.05）；与pcDNA3.1-NC组比较，过表达PERK组细胞增殖活性升高，细胞内E-cadherin荧光表达明显减弱（P < 0.05），E-cadherin蛋白表达下调（P < 0.05），促进细胞增殖、迁移与侵袭，抑制细胞凋亡（P < 0.05）；与sh-NC组比较，干扰PERK组细胞增殖活性降低，细胞内E-cadherin荧光表达则明显增强（P < 0.05），E-cadherin蛋白表达上调（P < 0.05），抑制细胞增殖、迁移与侵袭，促进细胞凋亡（P < 0.05）。结论 内质网应激PERK/eIF-2α在子宫内瘤变和CC组织中表达升高，抑制其表达能够降低CC肿瘤细胞增殖，从而减缓肿瘤进程。

Abstract

Objective To investigate the expression of the endoplasmic reticulum stress protein kinase R-like endoplasmic reticulum kinase (PERK)/eukaryotic initiation factor 2α (eIF-2α) signaling pathway in cervical intraepithelial neoplasia (CIN) and cervical carcinoma (CC), and its impact on tumor progression. Methods A total of 58 cervical biopsy specimens from patients diagnosed with cervical cancer (CC) treated at Nanjing Mingji Hospital from January 2022 to April 2023 were collected as the cervical cancer group. Concurrently, 52 cases of cervical intraepithelial neoplasia (CIN II and CIN III) were selected as the control group, and 34 cases of healthy uterine tissues from the same period were chosen as the normal group. Western blot was used to detect the expression of endoplasmic reticulum-mediated related proteins; immunohistochemistry was employed to assess the staining status of PERK/eIF-2α in CC tissues. HeLa cells were divided into the following groups: control group, pcDNA3.1-NC group, overexpressed PERK group, sh-NC group, and PERK interference group. Cell proliferation activity was detected using the CCK-8 assay, and the expression of epithelial cadherin (E-cadherin) was observed by immunofluorescence staining. Apoptosis and cell cycle distribution were analyzed by flow cytometry, and cell migration and invasion ability were evaluated by Transwell assay. Results With increasing CIN grade, the expression of PERK/eIF-2α increased (P < 0.05), and its expression in CC tissues was significantly higher than that in CIN tissues (P < 0.05). Compared with the pcDNA3.1-NC group, cells in the PERK overexpression group enhanced proliferation activity, markedly reduced intracellular E-cadherin fluorescence expression (P < 0.05), and downregulated E-cadherin protein expression (P < 0.05), accompanied by promotion of cell proliferation, migration, and invasion, as well as inhibition of apoptosis (P < 0.05). In contrast, compared with the sh-NC group, cells in the PERK interference group showed decreased proliferation activity, significantly increased intracellular E-cadherin fluorescence expression (P < 0.05), upregulated E-cadherin protein expression (P < 0.05), along with inhibition of cell proliferation, migration, invasion, and promotion of apoptosis (P < 0.05). Conclusion The expression of the endoplasmic reticulum stress-related PERK/eIF-2α pathway is upregulated in uterine intraepithelial neoplasia and CC tissues. Inhibition of its expression can reduce the tumor cell proliferation in CC, thereby slowing tumor progression.

关键词

宫颈癌 / 内质网应激 / 应激蛋白激酶R样内质网激酶 / 真核启动因子2α

Key words

Cervical cancer / Endoplasmic reticulum stress / Protein kinase R-like endoplasmic reticulum kinase / Eukaryotic initiation factor 2α

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基金资助

江苏省妇幼健康科研项目(F202178)