Characterization of a Polyclonal Antibody to Human Pituitary Tumor Transforming Gene 1 (PTTG1) Protein

Kakar, Sham S.; Chen, Leilei; Puri, Rashmi; Flynn, Shawn E; Jennes, Lothar

doi:10.1177/002215540104901207

Cited by 16 publications

(22 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2 and 3). PTTG is a commonly overexpressed gene in various types of cancer (5,6,(9)(10)(11)(12)(13)(14)(15)(16)(17)(18) and is believed to play an important role in augmenting tumor angiogenesis, metastasis and growth. Overexpression of PTTG has been shown to induce cellular transformation and to promote tumor development in nude mice.…”

Section: Discussionmentioning

confidence: 99%

“…Equal amounts of protein extracts (40 μg) were resolved on a 12% SDS-PAGE gel, and transferred onto a nitrocellulose membrane (Amersham Biosciences, Piscataway, NJ). Blots were probed with PTTG antiserum at a dilution of 1:1,500 as described previously (13). Immunoreactive proteins were visualized using the Enhanced Chemiluminescent Detection system (ECL) (Amersham Biosciences) according to instructions provided.…”

Section: Western Blot Analysismentioning

confidence: 99%

“…In contrast, PTTG is highly expressed in a variety of human primary tumors as well as tumor cell lines, including carcinoma of the ovary, testis, kidney, colon, thyroid, pituitary, liver, adrenal, and breast, and melanoma, leukemia, and lymphoma (5,6,(9)(10)(11)(12)(13)(14)(15)(16)(17)(18), suggesting that PTTG may be an important gene involved in human tumorigenesis. Furthermore, the expression levels of PTTG correlate positively with increased tumor invasiveness in hormone-producing human pituitary tumors (14) and the degree of malignancy, pathogenesis and/or progression of colorectal, thyroid and breast tumors (12,13,15). Recently, PTTG was identified as one of the signature genes among 8 associated with tumor metastasis that are up-regulated in human primary solid tumors (17).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Suppression of lung cancer with siRNA targeting PTTG

Kakar

Malik²

2006

Int J Oncol

View full text Add to dashboard Cite

Abstract. Pituitary transforming gene (PTTG) is frequently expressed at high levels in malignant tumors. We report high levels of expression of PTTG in various lung tumors and tumor-derived cell lines. For a better understanding of its role in maintaining the cancer phenotype, we used RNA interference (RNAi) directed against PTTG. Transfection of H1299 cells with PTTG siRNA duplex (5'-UGG GAG AUC UCA AGU UUC A-3') to a final concentration of 100 nmol/l resulted in almost complete depletion of PTTG mRNA within 24 and 48 h when compared to expression in untransfected cells or cells transfected with control siRNA. Western blot analysis showed nearly a 60% reduction in PTTG protein within 48 h of transfection. In phenotype analysis, we investigated the effect of PTTG siRNA on colony formation on soft agar, and tumor development in nude mice. Transfection of H1299 cells with PTTG siRNA duplex significantly reduced colony formation compared to untransfected cells or cells transfected with control siRNA. Mice injected with H1299 cells transfected with PTTG siRNA followed by injection of siRNA developed no tumors within two weeks of injection, but developed small tumors (67.85±45.87 mg) within 4 weeks of injection, whereas untransfected cells, or cells transfected with control siRNA developed large tumors (232.12±102.78 and 231.57±83.76 mg respectively). Suppression of PTTG as well as Ki67, bFGF and CD34 was observed in H1299 tumors treated with PTTG siRNA compared to untreated and control-treated siRNA in nude mice. In conclusion, decreasing PTTG expression through PTTG siRNA inhibits tumor growth both in vitro and in vivo. Future studies are needed to test whether PTTG expression can be efficiently depleted by siRNA expressed from a DNAbased expression vector combined with a specific-promoter, such that RNAi can specifically target PTTG in cancer cells without affecting normal cells.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Western Blot Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Suppression of lung cancer with siRNA targeting PTTG

Kakar

Malik²

2006

Int J Oncol

View full text Add to dashboard Cite

show abstract

“…In our previous studies using Northern blotting, reverse transcriptase/polymerase chain reaction (RT/PCR), and immunohistochemical analysis, we showed high levels of expression of PTTG mRNA and protein in ovarian tumors and tumor derived cell lines (8)(9)(10). In the present study, we used an ovarian cancer profiling array (Clontech, Palo Alto, CA) that included normalized cDNA from 10 different ovarian tumors and 10 corresponding normal ovarian tissues, along with negative and positive controls.…”

Section: Resultsmentioning

confidence: 98%

“…Initially cloned from the rat pituitary tumor (3), pituitary tumor transforming gene (PTTG), also known as securin, is an oncogene that is highly expressed in many tumors including pituitary (4,5), lung (6), colon, and ovarian tumors (7)(8)(9)(10). On the other hand, PTTG expression in normal tissue is very low except for the testis (8).…”

Section: Introductionmentioning

confidence: 99%

Small interfering RNA against PTTG: A novel therapy for ovarian cancer

El‐Naggar

Malik²,

Kakar³

2007

Int J Oncol

View full text Add to dashboard Cite

Abstract. Ovarian epithelial cancer is a significant cause of death among women, accounting for 5% of all female cancer-related fatalities. A lack of reliable detection methods and resistance to chemotherapy agents are considerable obstacles in the treatment of this cancer. Recently, high-level expression of the pituitary tumor transforming gene (PTTG) was found in a wide range of tumors, including ovarian cancers. Elevated PTTG levels were found to induce cellular transformation in vitro and tumor formation in nude mice. Therefore, we hypothesize a correlation exists between the levels of PTTG expression and tumorigenesis, and that downregulation of PTTG levels will result in the suppression of tumor growth. We used small interfering RNA (siRNA) to silence PTTG expression in human A2780 ovarian carcinoma cells and assessed the effect of PTTG silencing in tumor formation in vitro and in vivo. The siRNA directed against PTTG reduced its expression at both the mRNA and protein levels. A fifty percent reduction in cell proliferation was achieved in cells constitutively expressing PTTG siRNA compared to vector or control-siRNA transfected cells. Furthermore, colony formation in soft agar was reduced by 70% in PTTG siRNA stable cell lines. Using nude mice, we showed that animals injected with A2780 cells constitutively expressing PTTG-siRNA decreased the incidence of tumor development and tumor growth. Taken together, these results strongly suggest that PTTG may serve as an important molecular target for the discovery of new anticancer agents and treatment strategies.

show abstract