Abstract
Glucose regulated protein 78 (GRP78) is a type of molecular chaperone. It is a possible candidate protein that contributes to development of drug resistance. We first examined the involvement of GRP78 in chemotherapy-resistance in human ovarian cancer cell. The expression of GRP78 mRNA and protein were examined by RT-PCR and western blotting, respectively, in human ovarian cancer cells line (HO-8910). Sensitivity of HO-8910 to paclitaxel was determined with methyl thiazolyl tetrazolium (MTT). Suppression of GRP78 expression was performed using specific small-interfering RNA (siRNA) in HO-8910 cells, and cell apoptosis was assessed by flow cytometry. Statistical analysis was performed using the SPSS 15.0 statistical package. HO-8910 cells, with high basal levels of GRP78, exhibited low sensitivity to paclitaxel. The mRNA and protein levels of GRP78 were dramatically decreased at 24h, 48h and 72h after transfection and the sensitivity to paclitaxel was increased when the GRP78 gene was disturbed by specific siRNA transfection. The results suggested that high GRP78 expression might be one of the molecular mechanisms causing resistance to paclitaxel, and therefore siRNA of GRP78 may be useful in tumor-specific gene therapy for ovarian cancer.
Highlights
Ovarian carcinoma (OC) is the most lethal of the gynaecological malignancies and it is the fourth most common cause of cancer-related death in women (Sevim et al, 2014)
The results suggested that high Glucose regulated protein 78 (GRP78) expression might be one of the molecular mechanisms causing resistance to paclitaxel, and small-interfering RNA (siRNA) of GRP78 may be useful in tumor-specific gene therapy for ovarian cancer
To deregulate the basal GRP78 mRNA expression, siRNA targeted to GRP78 was designed and transfected into HO-8910 cells and nonspecific siRNA was designed and used for observing the non-specific effect of siRNA transfection
Summary
Ovarian carcinoma (OC) is the most lethal of the gynaecological malignancies and it is the fourth most common cause of cancer-related death in women (Sevim et al, 2014). We first examined the involvement of GRP78 in chemotherapy-resistance in human ovarian cancer cell. Materials and Methods: The expression of GRP78 mRNA and protein were examined by RT-PCR and western blotting, respectively, in human ovarian cancer cells line (HO-8910). Suppression of GRP78 expression was performed using specific small-interfering RNA (siRNA) in HO-8910 cells, and cell apoptosis was assessed by flow cytometry. Results: HO-8910 cells, with high basal levels of GRP78, exhibited low sensitivity to paclitaxel. The mRNA and protein levels of GRP78 were dramatically decreased at 24h, 48h and 72h after transfection and the sensitivity to paclitaxel was increased when the GRP78 gene was disturbed by specific siRNA transfection. Conclusions: The results suggested that high GRP78 expression might be one of the molecular mechanisms causing resistance to paclitaxel, and siRNA of GRP78 may be useful in tumor-specific gene therapy for ovarian cancer
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