title: Gene transfections with p53 and p21 inhibit cell proliferation, collagen type I, leukemia inhibitory factor, and tumor necrosis factor-alpha expression in leiomyoma cells.
abstract: OBJECTIVE
  To transfect the p53 and p21 gene into the leiomyoma cells isolated from patients and observe their influence on the cell proliferation, leukemia inhibitory factor production, and gene expression of collagen type I as well as tumor necrosis factor-alpha (TNF-alpha) of cultured cells.
  
  
  METHODS
  Prospective study.
  
  
  METHODS
  An assisted reproductive technology (ART) and genetic unit of a medical center.
  
  
  METHODS
  Leiomyoma cells isolated from leiomyoma tissue of 12 patients were divided into three groups: [1]. vehicle DNA, [2]. p53 gene, and [3]. p21 gene transfections.
  
  
  METHODS
  The pcDNA3.1 was used as vector to carry p53 and p21 genes for transfer. After gene transfection, RNAs of the leiomyoma cells were extracted for further analyses of gene expression.
  
  
  METHODS
  Relative cell numbers were determined by 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay. The leukemia inhibitory factor (LIF) concentration was determined with ELISA. Gene expressions of collagen type I and TNF-alpha were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Gene expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control. The cell proliferation, LIF production, as well as gene expressions of collagen type I and TNF-alpha in each group were compared.
  
  
  RESULTS
  Relative cell numbers (%)/LIF production (in picograms per milliliter) in each group were: [1]. 100/58, [2]. 71/43, and [3]. 106/65. The ratios of gene expression of collagen type I/TNF-alpha with GAPDH in each group were: [1]. 1.64/0.335, [2]. 1.25/0.434, and [3]. 1.77/0.234.
  
  
  CONCLUSIONS
  Transfection with p53 significantly inhibits proliferation of leiomyoma cells and decreases collagen type I gene expression and LIF production. The p21 transfection inhibits TNF-alpha gene expression.
authors.affiliation: Department of Obstetrics and Gynecology, China Medical College Hospital, Taichung, Taiwan.
authors.forename: Yao-Yuan
authors.initials: YY
authors.lastname: Hsieh
authors.forename: Chi-Chen
authors.initials: CC
authors.lastname: Chang
authors.forename: Chih-Wei
authors.initials: CW
authors.lastname: Hsu
authors.forename: Chich-Sheng
authors.initials: CS
authors.lastname: Lin
mesh_terms: D002455:Cell Division; D024042:Collagen Type I / Q000037:antagonists & inhibitors*; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics*; D005260:Female; D016158:Genes, p53; D006801:Humans; D007889:Leiomyoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D053605:Leukemia Inhibitory Factor; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors*; D014594:Uterine Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D000093242:Interleukin-6 Inhibitors
chemical_list: D024042:Collagen Type I; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D053605:Leukemia Inhibitory Factor; D014409:Tumor Necrosis Factor-alpha; D000093242:Interleukin-6 Inhibitors; C493956:CDKN1A protein, human; C507837:LIF protein, human