Summary
The reversing effect of wild-type PTEN gene on resistance of C13K cells to cisplatin and its inhibitory effect on the phosphorylation of protein kinase B (AKT) were studied. The expression of PTEN mRNA and protein in OV2008 cells and C13K cells were semi-quantitatively detected by using RT-PCR and Western blotting. Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine2000. The expression of PTEN mRNA was monitored by RT-PCR and the expression of PTEN, Akt, p-Akt protein were analyzed by Western blotting in PTEN-transfected and non-transfected C13K cells. Proliferation and chemosensitivity of cells to DDP were measured by MTT, and cell apoptosis was detected by flow cytometry after treatment with cisplatin. The expression of PTEN mRNA and protein in OV2008 cells were significantly higher than those in C13K cells. After transfection with PTEN gene for 48 h, the expression of PTEN mRNA and protein in C13K cells were 2.04 ± 0.10, 0.94 ± 0.04 respectively and the expression of p-Akt protein (0.94 ± 0.07) was lower than those in control groups (1.68 ± 0.14, 1.66 ± 0.10) (P < 0.05). The IC50 of DDP to C13K cells transfected with PTEN (7.2 ± 0.3 μmol/L) was obviously lower than those of empty-vector transfected cells and non-transfected cells (12.7 ± 0.4 μmol/l, 13.0 ± 0.3 μmol/L) (P<0.05). The apopototis ratio of wild-type PTEN-transfected, empty vector transfected and non-transfected C13K cells were (41.65 ± 0.87)%, (18.61 ± 0.70)% and (15.28 ± 0.80)% respectively, and the difference was statistically significant (P<0.05). PTEN gene plays an important role in ovarian cancer multidrug resistance. Transfection of PTEN could increase the expression of PTEN and restore drug sensitivity to cisplatin in human ovarian cancer cell line C13K with multidrug-resistance by decreasing the expression of p-Akt.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Sehondorf T, Becker M, Gohring U J et al. Interaction of cisplatin, paclitaxel and adriamycin with the tumor suppressor PTEN. Anticancer Drugs, 2001(10):797–800
Tamura M, Cu J, Takino T et al. Tumor suppressor PTEN inhibition of cell invasion, migration and growth: differential involvement of focal adhesion kinase and pl30Cas. Cancer Res, 1999,59(1):442–449
Li J, Yen C, Liaw D et al. PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostatecancer. Science, 1997,275(5308):1943–1947
Di-Cristofano A, Pandolfi P. The multiple roles of PTEN in tumor suppression. Cell, 2000,100(4):387–390
Cantley L C, Neel B G. New insights into tumour suppression: PTEN suppresses tumour formation by restraining the phosphoinositide 3-kinase/Akt pathway. Proc Natl Acad Sci USA, 1999,96(8):4240–4245
Tanaka M, Grossman H B. In vivo gene therapy of human bladder cancer with PTEN suppresses tumor growth, downregulates phosphorylated Akt, and increases sensitivity to doxorubicin. Gene Ther, 2003,10(19):1636–1642
Lee E S, Choi E J, Jn C et al. Activation of P I3K/A k t pathway by PTEN reduction and P IK3CA mRNA amplification contributes to cisplatin resistance in an ovarian cancer cell line. Gynecol Oncol, 2005,97(1):26–34
Nagata Y, Lan K H, Zhou X et al. PTEN activation contributes to tumor inhibition by trastuzumab, and loss of PTEN predicts trastuzumab resistance in patients. Cancer Cell, 2004,6(8):117–127
Zhou M, Gu L, Findley H et al. PTEN reverses mdm2-mediated chemotherapy resistance by interacting with p53 in actue lymphoblastic leukemia cells. Cancer Res, 2003,63(10):6357–6362
Yan X, Fraser M, Qiu Q et al. Over-expression of PTEN sensitizes human ovarian cancer cells to cisplatin-induced apoptosis in a p-53-depentent manner. Gynecol Oncol, 2006,102(2):348–355
Vivanco I, Sawyers C L. The phosphatidylinositol 3-kinase Akt pathway in human cancer. Nat Rev Cancer, 2002,2(7):489–501
Gagnon V, Mathieu I, Sexton E et al. AKT involvement in cisplatin chemoresistance of human uterine cancer cells. Gynecol Oncol, 2004,94(3):785–795
Author information
Authors and Affiliations
Additional information
This project was supported by a grant from the National Natural Sciences Foundation of China (No. 30571950) and National Key Basic Research Program Foundation (NO.2002CB513107).
Rights and permissions
About this article
Cite this article
Wu, H., Weng, D., Xing, H. et al. Reversal of multidrug resistance and inhibition of phosphorylation of AKT in human ovarian cancer cell line by wild-type PTEN gene. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 27, 713–716 (2007). https://doi.org/10.1007/s11596-007-0625-9
Received:
Issue Date:
DOI: https://doi.org/10.1007/s11596-007-0625-9