Epigenetic modification-mediated inhibition of SOSTDC1 expression promotes malignant biological behaviors in cervical cancer cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究含硬化蛋白域蛋白1（SOSTDC1）对宫颈癌细胞恶性生物学行为的调控及其分子机制。方法：收集2020年8月至2022年5月间在福建省肿瘤医院活检或手术切除的53例宫颈癌组织和相应的癌旁组织标本，免疫组化法检测SOSTDC1蛋白在宫颈癌组织及相应癌旁组织中的表达，qPCR法检测正常宫颈细胞、宫颈癌细胞中SOSTDC1 mRNA表达；将SOSTDC1过表达慢病毒（OE-sostdc1）和对照空病毒（NC）感染宫颈癌细胞SiHa及CaSki，将其分为SiHa-OE-sostdc1、SiHa-NC、CaSki-OE-sostdc1、CaSki-NC组，采用WST-1法、细胞集落形成实验、Transwell实验和WB法检测转染各组SiHa及CaSki细胞的增殖、集落形成、迁移和侵袭能力和BMP、Wnt/β-catenin信号途径相关蛋白及上皮-间充质转化（EMT）相关蛋白的表达。用DNA甲基化酶抑制剂5-氮杂2'-脱氧胞苷（5'-Aza-CdR）处理宫颈癌细胞后采用qPCR和WB法检测SOSTDC1 mRNA及蛋白的表达变化，用甲基化特异性PCR（MSP）检测5例配对宫颈癌组织与癌旁组织中SOSTDC1基因启动子区甲基化水平，同时qPCR检测其SOSTDC1 mRNA水平。结果：与癌旁组织比较，SOSTDC1蛋白在宫颈癌组织中呈低表达（P<0.01），且与淋巴结转移与FIGO分期有关联（均P<0.05）；与正常宫颈HUCEC细胞比较，SOSTDC1 mRNA在宫颈癌C33A、HeLa、SiHa、CaSki细胞中均呈低表达（均P<0.01）。过表达SOSTDC1显著抑制SiHa及CaSki细胞的增殖、迁移和侵袭能力（均P<0.05）。WB法结果检测显示，过表达SOSTDC1显著抑制SiHa及CaSki细胞中磷酸化Smad、Dvl2/3、β-catenin、VIM、N-cadherin、Snail蛋白的表达（均P<0.05），5'-Aza-CdR处理后的SiHa及CaSki细胞中SOSTDC1 mRNA和蛋白水平均显著增加（均P<0.05），MSP检测结果显示，相较于癌旁组织，宫颈癌组织中SOSTDC1基因启动子区呈高度甲基化，且SOSTDC1 mRNA水平降低（P<0.01）。结论：SOSTDC1在宫颈癌组织中呈低表达且与肿瘤的恶性进展关联，其表达下调与其基因启动子区高度甲基化有关，过表达SOSTDC1可能通过阻断BMP及Wnt/β-catenin信号通路从而抑制SiHa、CaSki细胞的增殖、侵袭和迁移能力。

Id1 and Id3 synergistically influence invasion and migration of colon cancer SW620 cells by inducing EMT / 中国肿瘤生物治疗杂志

@# Objective: To investigate whether inhibitor of differentiation 1 gene (Id1) and Id3 gene can synergistically promote epithelial-mesenchymal transition (EMT), invasion and migration of colon cancer SW620 cells and to explore its underlying mechanisms. Methods: The SW620 cell strain with Idl or Id3 gene knockdown and the SW620 cell strain with Id1/Id3 gene double-knockdown were constructed by lentiviral vectors transfection. The SW620 cells were divided into four groups, which included SW620-Sh-Id1 group (transfected with shRNA-Id1), SW620-Sh-Id3 group (transfected with shRNA-Id3), SW620-Sh-Id1-Id3 group (transfected with shRNAId1 plus shRNA-Id3) and SW620-NC group (transfected with negative lentivirus). The efficiency of knockdown was detected by Realtime qPCR and Western blotting. The influence of stable knockdown of Idl or Id3 on cell morphological change was observed under a microscope. The changes of migration and invasion abilities of the SW620 cells were determined by wound healing assay and Transwell assay. EMT, invasion and migration related proteins were measured by Western blotting. Results: The SW620 cell strains with Idl and/or Id3 gene knockdown were successfully constructed. Idl and Id3 knockdown induced the epithelial-like to the mesenchymanl-like transformation of SW620 cells. (1) Compared with the control group, the invasion and migration abilities of the SW620 cells were significantly decreased in the SW620-Sh-Id1 group and SW620-Sh-Id3 group (all P<0.05). (2) Meanwhile, the invasion and migration abilities in the SW620-Sh-Id1-Id3 group were obviously weaker than the SW620-Sh-Id1 group and SW620-Sh-Id3 group (all P<0.05). (3) Compared with the control group, the SW620-Sh-Id1 group and SW620-Sh-Id3 group had a reduction in the protein expressions of βcatenin, snail1 and MMP2, and an increase in the protein expressions of E-cadherin and TIMP2 (all P<0.05). (4) Compared with the SW620-Sh-Id1 group and SW620-Sh-Id3 group , the protein expressions of β-catenin, snail1 and MMP2 were reduced, and the protein expressions of E-cadherin and TIMP2 were increased in the SW620-Sh-Id1-Id3 group (all P<0.05). Conclusion: Id1 and Id3 could synergistically influence invasion and migration of SW620 cells, possibly through inducing EMT.