Preliminary study of the prohibitin protein and paclitaxel resistance in ovarian cancer.
10.3969/j.issn.1672-7347.2012.12.007
- Author:
Juan TANG
1
;
Lanqin CAO
;
Hong YI
;
Can'e TANG
Author Information
1. Department of Gynecology and Obstetrics, Central South University, Changsha, China.
- Publication Type:Journal Article
- MeSH:
Antineoplastic Agents, Phytogenic;
pharmacology;
Drug Resistance, Neoplasm;
genetics;
Female;
Humans;
Ovarian Neoplasms;
genetics;
metabolism;
pathology;
Paclitaxel;
pharmacology;
RNA Interference;
RNA, Messenger;
genetics;
metabolism;
RNA, Small Interfering;
genetics;
Repressor Proteins;
genetics;
metabolism;
Transfection;
Tumor Cells, Cultured
- From:
Journal of Central South University(Medical Sciences)
2012;37(12):1221-1227
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To determine the effect of RNA interference with transferred pshRNA/PHB on the biological characteristics of paclitaxel-resistant ovarian cancer cell lines.
METHODS:Western blot and real time-PCR were used to assay the expression of PHB protein and mRNA in SKOV3/Taxol-25 and SKOV3 cell lines. The SKOV3/Taxol-25 cell lines were transiently transfected by 3 target-specific small hairpin RNA (shRNA) interference fragments with fluorescent protein named the pshRNA427/PHB1, pshRNA248/PHB2, and pshRNA136/PHB3. The empty plasmid transfection via vehicle Lipofectamine2000 served as a negative control. The expression levels of PHB protein and mRNA were detected by Western blot and real time-PCR after the transfection for 48 h. The silence effect of PHB1 and PHB3 groups was obvious. PHB1, PHB3, and the negative control groups were used for the following experiments. MTT and flow cytometry assay were used to test the cell proliferation, IC50 of paclitaxel, and cell apoptosis in the 3 groups.
RESULTS:The expression levels of PHB protein and mRNA (2(-ΔΔCt)) were significantly higher in SKOV3/Taxol-25 cell line than those in SKOV3 cell line (P<0.05). The expression levels of PHB protein and mRNA were significantly lower in the PHB1 and PHB3 groups than those in the negative control group (P<0.05). The cell proliferations in the PHB1 and PHB3 groups were obviously slower than those in the negative control group after transfection for 48 h and 72 h (P<0.05). The IC50 of paclitaxel in the PHB1 and PHB3 groups significantly decreased after transfection for 72 h compared with the negative control group(P<0.05). The cell apoptotic rate in the PHB1 and PHB3 groups significantly increased after transfection for 48 h compared with the negative control group (P<0.05).
CONCLUSION:The shRNA/PHB can effectively suppress the expression of PHB gene in paclitaxel-resistant ovarian cancer cell lines. The cell proliferation in paclitaxel-resistant cell lines with removed PHB gene is significantly reduced. The apoptotic rate and the paclitaxel sensitivity of resistant cell lines with removed PHB gene are significantly increased. PHB gene is related to paclitaxel-resistance and interfering PHB gene expression may reduce paclitaxel resistance in ovarian cancer.
