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下调 SEMA4C 通过抑制转化生长因子-β1(TGF-β1)诱导的 Hela 细胞 p38 丝裂原活化蛋白激酶(MAPK)激活抑制宫颈癌上皮间质转化(EMT)和侵袭转移。

Downregulation of SEMA4C Inhibit Epithelial-Mesenchymal Transition (EMT) and the Invasion and Metastasis of Cervical Cancer Cells via Inhibiting Transforming Growth Factor-beta 1 (TGF-β1)-Induced Hela cells p38 Mitogen-Activated Protein Kinase (MAPK) Activation.

机构信息

Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China (mainland).

Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China (mainland).

出版信息

Med Sci Monit. 2020 Jan 17;26:e918123. doi: 10.12659/MSM.918123.

Abstract

BACKGROUND Epithelial-mesenchymal transition (EMT) plays a key role in promoting invasion and metastasis of tumor cells. SEMA4C can regulate the generation of transforming growth factor-beta 1 (TGF-ß1)-induced EMT in cervical cancer. This study investigated the relationship between the regulation of SEMA4C on TGF-ß1-induced p38 mitogen-activated protein kinase (MAPK) activation and invasion and metastasis of cervical cancer. MATERIAL AND METHODS Hela-shSEMA4C cell line was established and the success of transfection was confirmed with fluorescence intensity. Cell experiments were divided into 2 groups. Group 1 was Hela, Hela-shNC, and Hela-shSEMA4C; and Group 2 was Hela, Hela-shNC, Hela-shSEMA4C, Hela+TGF-ß1, Hela-shNC+TGF-ß1, and Hela-shSEMA4C+TGF-ß1. Group 1 was detected for SEMA4C mRNA expression by real-time polymerase chain reaction (RT-PCR), cell viability by Cell Counting Kit-8 (CCK-8), F-actin fluorescence intensity by immunofluorescence, cell migration by scratch test, and cell invasion by invasion test. Group 2 was analyzed for E-cadherin fluorescence intensity by immunofluorescence, human fibronectin (FN) content by enzyme-linked immunosorbent assay (ELISA), and SEMA4C, E-cadherin and p-p38 expressions by Western blot. RESULTS For Group 1, compared with Hela and Hela-shNC subgroups, the SEMA4C mRNA expression, cell viability, F-actin fluorescence intensity, cell migration and invasion ability in the Hela-shSEMA4C subgroup were significantly decreased (P<0.05). For Group 2, compared with Hela and Hela-shNC subgroups, the E-cadherin expression and fluorescence intensity in the Hela-shSEMA4C subgroup were significantly increased (P<0.01), while the FN content, SEMA4C, and p-p38 MAPK expressions were significantly decreased (P<0.01). Compared with Hela-shNC+TGF-ß1 and Hela+TGF-ß1 subgroups, the E-cadherin expression and fluorescence intensity in the Hela-shSEMA4C+TGF-ß1 subgroup were significantly increased (P<0.01), while the FN content, SEMA4C and p-p38 expressions were significantly decreased (P<0.01). CONCLUSIONS Downregulation of SEMA4C can inhibit EMT and the invasion and metastasis of cervical cancer cells via inhibiting TGF-ß1-induced Hela cells p38 MAPK activation.

摘要

背景

上皮-间充质转化(EMT)在促进肿瘤细胞侵袭和转移中起关键作用。SEMA4C 可调节宫颈癌中转化生长因子-β1(TGF-β1)诱导的 EMT 的产生。本研究探讨了 SEMA4C 对 TGF-β1 诱导的 p38 丝裂原活化蛋白激酶(MAPK)激活与宫颈癌侵袭和转移的关系。

材料与方法

建立了 Hela-shSEMA4C 细胞系,并通过荧光强度确认转染的成功。细胞实验分为 2 组。第 1 组为 Hela、Hela-shNC 和 Hela-shSEMA4C;第 2 组为 Hela、Hela-shNC、Hela-shSEMA4C、Hela+TGF-β1、Hela-shNC+TGF-β1 和 Hela-shSEMA4C+TGF-β1。第 1 组通过实时聚合酶链反应(RT-PCR)检测 SEMA4C mRNA 表达、细胞计数试剂盒-8(CCK-8)检测细胞活力、免疫荧光检测 F-肌动蛋白荧光强度、划痕试验检测细胞迁移、侵袭试验检测细胞侵袭。第 2 组通过免疫荧光检测 E-钙黏蛋白荧光强度、酶联免疫吸附试验(ELISA)检测人纤维连接蛋白(FN)含量、Western blot 检测 SEMA4C、E-钙黏蛋白和 p-p38 表达。

结果

第 1 组与 Hela 和 Hela-shNC 亚组相比,Hela-shSEMA4C 亚组的 SEMA4C mRNA 表达、细胞活力、F-肌动蛋白荧光强度、细胞迁移和侵袭能力显著降低(P<0.05)。第 2 组与 Hela 和 Hela-shNC 亚组相比,Hela-shSEMA4C 亚组的 E-钙黏蛋白表达和荧光强度显著增加(P<0.01),而 FN 含量、SEMA4C 和 p-p38 MAPK 表达显著降低(P<0.01)。与 Hela-shNC+TGF-β1 和 Hela+TGF-β1 亚组相比,Hela-shSEMA4C+TGF-β1 亚组的 E-钙黏蛋白表达和荧光强度显著增加(P<0.01),而 FN 含量、SEMA4C 和 p-p38 表达显著降低(P<0.01)。

结论

下调 SEMA4C 通过抑制 TGF-β1 诱导的 Hela 细胞 p38 MAPK 激活,抑制 EMT 及宫颈癌细胞的侵袭和转移。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b9a6/6986213/dcc6296c7951/medscimonit-26-e918123_g001.jpg

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