Gene transfections with p53 and p21 inhibit cell proliferation, collagen type I, leukemia inhibitory factor, and tumor necrosis factor-alpha expression in leiomyoma cells.
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.
Hsieh
Yao-Yuan
YY
Department of Obstetrics and Gynecology, China Medical College Hospital, Taichung, Taiwan.
Chang
Chi-Chen
CC
Hsu
Chih-Wei
CW
Lin
Chich-Sheng
CS
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
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