NDLI: Effect of cyclin-dependent kinase 7 silencing on cisplatin sensitivity in endometrial carcinoma cells.

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Effect of cyclin-dependent kinase 7 silencing on cisplatin sensitivity in endometrial carcinoma cells.

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Effect of cyclin-dependent kinase 7 silencing on cisplatin sensitivity in endometrial carcinoma cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Hao, Quan Yu, Hu Liu, Wen-Xin Liu, Xiang-Yu Chen, Ying
Description	Author Affiliation: Liu WX ( Department of Gynecologic Neoplasms, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, P.R. China.); Liu XY ( Department of Gynecologic Neoplasms, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, P.R. China.); Yu H ( Department of Gynecologic Neoplasms, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, P.R. China.); Chen Y ( Department of Gynecologic Neoplasms, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, P.R. China.); Hao Q ( Department of Gynecologic Neoplasms, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, P.R. China.)
Abstract	The aim of the present study was to determine the effect of cyclindependent kinase 7 (CDK7) silencing on the sensitivity of the HEC1A endometrial carcinoma cell line to cisplatin [cisdichlorodiammineplatinum (II), or DDP]. Four CDK7 siRNA fragments were designed and synthesized based on the gene sequence of CDK7 and transfected into HEC1A cells. The RNA interference of the fragments was confirmed by semiquantitative polymerase chain reaction (PCR) and western blot analyses. The CDK7423 siRNA fragment exhibited the most marked silencing of CDK7 (>70%), and was chosen for the subsequent experiments in HEC1A endometrial carcinoma cells. The sensitivity of the cells to a chemotherapeutic agent (cisplatin) was determined before and after transfection of the siRNA, using a MTT cytotoxicity assay, flow cytometry and Hoechst/propidium iodide (PI) doublestaining immunofluorescence microscopy. The results of the MTT cytotoxicity assay showed that the half maximal inhibitory concentration of cisplatin was reduced from 45.12 µg/ml to 3.200 µg/ml following the inhibition of CDK7 expression levels, indicating a significantly increased cytotoxicity in the treated cells (P<0.05). The flow cytometry analysis showed that the mean rate of apoptosis in the CDK7 lowexpression group was 37.57%, which was significantly higher than the rate in the parental cells (11.66%) (P<0.05). Hoechst/PI coimmunofluorescence microscopy revealed that the number of apoptotic bodies in the CDK7 lowexpression HEC1A cells was significantly increased as compared with the parental cells. Downregulation of CDK7 expression levels in HEC1A endometrial carcinoma cells via the transfection of CDK7 siRNA may significantly enhance cancer cell sensitivity to cisplatin chemotherapy and increasing apoptosis. CDK7 is a novel promising treatment for endometrial carcinoma that requires further indepth study.
ISSN	17912997
e-ISSN	17913004
DOI	10.3892/mmr.2014.2980
Journal	Molecular Medicine Reports
Issue Number	3
Volume Number	11
Language	English
Publisher	Spandidos Publications
Publisher Date	2015-03-01
Publisher Place	Greece
Access Restriction	Open
Subject Keyword	Antineoplastic Agents Pharmacology Cisplatin Cyclin-dependent Kinases Genetics Drug Resistance, Neoplasm Gene Silencing Ovarian Neoplasms Apoptosis Drug Effects Cell Cycle Cell Line, Tumor Gene Expression Rna Interference Rna, Small Interfering Transfection Discipline Molecular Biology
Content Type	Text
Resource Type	Article
Subject	Genetics Biochemistry Molecular Biology Cancer Research Molecular Medicine Oncology

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