Department of Gynecology, the First Affiliated Hospital of Bengbu Medical College, 287 Changhuai Road, Bengbu, 233004, Anhui, China.
J Mol Histol. 2023 Oct;54(5):489-498. doi: 10.1007/s10735-023-10145-6. Epub 2023 Aug 24.
Cervical cancer (CC) is the second most common type of cancer in women, and presents a serious threat to public health. We aimed to investigate the regulatory impacts of CDGSH iron-sulfur domain-containing protein 2 (CISD2) in CC and to discuss its relationship with E2F transcription factor 7 (E2F7). With the employment of real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and western blot, the expression of CISD2 and E2F7 in SiHa cells before or after transfection was estimated. Cell counting kit-8 (CCK-8) assay, Terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay, wound healing and transwell were used to detect the proliferation, apoptosis, migration and invasion of SiHa cells. The activity of CISD2 was detected using luciferase report assay and chromatin immunoprecipitation (ChIP) assay was used to confirm the binding of E2F7 and CISD2 promoter. The contents of proliferation- and apoptosis-related proteins were detected using western blot. Results revealed that CISD2 expression was greatly enhanced in CC cell lines. CISD2 depletion inhibited the proliferation, migration and invasion of SiHa cells but promoted the cell apoptosis. It was also found that E2F7 was remarkably elevated in SiHa cells. According to JASPAR database, the binding sites of E2F7 and CISD2 were predicted and ChIP confirmed the binding of E2F7 and CISD2 promoter. Results obtained from luciferase report assay indicated that E2F7 overexpression increased the activity of CISD2 promoter region. Furthermore, further functional experiments demonstrated that the impacts of E2F7 interference on the proliferation, migration, invasion and apoptosis of SiHa cells were reversed by CISD2 overexpression. In summary, CISD2 silence could alleviate the malignant progression of CC and could be transcribed by E2F7.
宫颈癌（CC）是女性中第二常见的癌症类型，对公众健康构成严重威胁。本研究旨在探讨钙-结合二硫簇结构域蛋白 2（CISD2）在 CC 中的调控作用，并探讨其与转录因子 E2F7 的关系。采用实时逆转录-聚合酶链反应（RT-qPCR）和蛋白质印迹法检测 SiHa 细胞转染前后 CISD2 和 E2F7 的表达。通过细胞计数试剂盒-8（CCK-8）检测、末端脱氧核苷酸转移酶（TdT）dUTP 末端标记（TUNEL）检测、划痕愈合和 Transwell 检测 SiHa 细胞的增殖、凋亡、迁移和侵袭。通过荧光素酶报告基因检测来检测 CISD2 的活性，并通过染色质免疫沉淀（ChIP）实验来确认 E2F7 和 CISD2 启动子的结合。通过蛋白质印迹法检测增殖和凋亡相关蛋白的含量。结果表明，CISD2 在 CC 细胞系中的表达显著增加。CISD2 耗竭抑制 SiHa 细胞的增殖、迁移和侵袭，但促进细胞凋亡。此外，还发现 E2F7 在 SiHa 细胞中显著上调。根据 JASPAR 数据库，预测了 E2F7 和 CISD2 的结合位点，ChIP 实验证实了 E2F7 与 CISD2 启动子的结合。荧光素酶报告基因检测结果表明，E2F7 过表达增加了 CISD2 启动子区域的活性。此外，进一步的功能实验表明，E2F7 干扰对 SiHa 细胞增殖、迁移、侵袭和凋亡的影响可被 CISD2 过表达逆转。综上所述，CISD2 沉默可减轻 CC 的恶性进展，可被 E2F7 转录。
