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. 2009 Oct;29(10):695-703.
doi: 10.1089/jir.2009.0003.

Down-regulation of GRIM-19 expression is associated with hyperactivation of STAT3-induced gene expression and tumor growth in human cervical cancers

Ying Zhou  1 , Min Li, Ying Wei, Dingqing Feng, Cheng Peng, Haiyan Weng, Yang Ma, Liang Bao, Shreeram Nallar, Sudhakar Kalakonda, Weihua Xiao, Dhananjaya V Kalvakolanu, Bin Ling
Affiliations

Down-regulation of GRIM-19 expression is associated with hyperactivation of STAT3-induced gene expression and tumor growth in human cervical cancers

Ying Zhou et al. J Interferon Cytokine Res. 2009 Oct.

Abstract

Cervical cancer is the most common malignant disease responsible for the deaths of a large number of women in the developing world. Although certain strains of human papillomavirus (HPV) have been identified as the cause of this disease, events that lead to formation of malignant tumors are not fully clear. STAT3 is a major oncogenic transcription factor involved in the development and progression of a number of human tumors. However, the mechanisms that result in loss of control over STAT3 activity are not understood. Gene associated with Retinoid-Interferon-induced Mortality-19 (GRIM-19) is a tumor-suppressive protein identified using a genetic technique in the interferon/retinoid-induced cell death pathway. Here, we show that reduction in GRIM-19 protein levels occur in a number of primary human cervical cancers. Consequently, these tumors tend to express a high basal level of STAT3 and its downstream target genes. More importantly, using a surrogate model, we show that restoration of GRIM-19 levels reestablishes the control over STAT3-dependent gene expression and tumor growth in vivo. GRIM-19 suppressed the expression of tumor invasion- and angiogenesis-associated factors to limit tumor growth. This study identifies another major novel molecular pathway inactivated during the development of human cervical cancer.

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Figures

FIG. 1.
FIG. 1.
Immunohistochemical analysis of normal cervical tissue and cervical cancer with gene associated with retinoid-interferon-induced mortality-19 (GRIM-19) antibodies. A representative picture of 15 independent normal samples and 20 tumor tissues are shown. An intense staining of GRIM-19 in normal cervical tissues is significantly suppressed in cervical tumors (200×). Photomicrographs on the right show a higher magnification (800×) of the boxed areas.
FIG. 2.
FIG. 2.
Down-regulation of GRIM-19 and up-regulation of STAT3 in cervical cancers. (A) RT-PCR analysis of the expression of GRIM-19, STAT3, Cyclin B1, and Bcl2-L1 in normal cervical tissue and cervical cancer tissue. (B) Equal amounts of total cellular extracts (50 μg) from the normal and tumor samples were resolved by SDS-PAGE and immunoblotted with anti-GRIM-19 and anti-STAT3 antibodies. Individual patient samples are indicated on top; N: normal; T: tumor.
FIG. 3.
FIG. 3.
Overexpression of GRIM-19 suppresses HeLa cell proliferation. (A) RT-PCR analysis of the expression of GRIM-19 in HeLa/Control and HeLa/GRIM-19 cells. (B) Western blot analysis of the expression of GRIM-19 and STAT3 in HeLa/Control and HeLa/GRIM-19 cells. (C) An MTT assay was performed in the indicated cells. MTT reduction, due to cell growth, converts the colorless dye into a chromogen, whose levels were quantified at 570 nm using microplate reader. Each data point represents mean ± SE of 8 samples. Cells were treated with a combination of 1 μM retinoic acid and 1,000 U of interferon-β and were indicated with R/I.
FIG. 4.
FIG. 4.
Restoration of GRIM-19 inhibits the tumor growth. (A) Representative mice bearing HeLa/Control and HeLa/GRIM-19 tumors and the tumors developed in them are shown in the photographs. Tumors harvested on day 42 are shown. (B) Antitumor effects of GRIM-19 in vivo. Tumor volumes were measured over a period of 6 weeks. The tumor volume differences between the GRIM-19 and control group is highly significant (*P < 0.01). (C) Western blot analysis of the expression of GRIM-19, STAT3, p-STAT3, and Survivin in the tumor tissues. (D) RT-PCR analysis of the expression of GRIM-19 and STAT3, Cyclin B1, and Bcl2-L1 in the tumor tissues.
FIG. 5.
FIG. 5.
GRIM-19 suppresses tumor angiogenesis and tumor invasion-associated gene expression. (A) Western blot analysis of the expression of tumor angiogenesis and metastasis-associated proteins (MMP-2 and MMP-9) and VEGF. A representative blot from several individual tumors (n = 4) is shown. (B) GRIM-19 suppresses tumor angiogenesis. Blood vessels were stained with CD31 antibodies. Note the loss of blood vessels in tumors with GRIM-19 (200×). Circled inserts show a magnified view (800×). Brown color deposits indicate positive staining for blood vessels. (C) Quantification of blood vessels in the tumors. Multiple regions (n = 5) from different tumors (n = 4) were quantified after staining with CD31. (D) TUNEL staining of tumors from HeLa/Control and HeLa/GRIM-19 (400×). Circled inserts show a magnified view (800×) of tumor apoptosis. (E) Quantification of apoptotic cells in tumors. Multiple regions (n = 5) from different tumors (n = 4) were quantified after TUNEL staining.
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References

    1. Alchanati I. Nallar SC. Sun P. Gao L. Hu J. Stein A. Yakirevich E. Konforty D. Alroy I. Zhao X. Reddy SP. Resnick MB. Kalvakolanu DV. A proteomic analysis reveals the loss of expression of the cell death regulatory gene GRIM-19 in human renal cell carcinomas. Oncogene. 2006;25(54):7138–7147. - PubMed
    1. Braun K. Ehemann V. Waldeck W. Pipkorn R. Corban-Wilhelm H. Jenne J. Gissmann L. Debus J. HPV18 E6 and E7 genes affect cell cycle, pRB and p53 of cervical tumor cells and represent prominent candidates for intervention by use peptide nucleic acids (PNAs) Cancer Lett. 2004;209(1):37–49. - PubMed
    1. Bromberg JF. Horvath CM. Besser D. Lathem WW. Darnell JE., Jr Stat3 activation is required for cellular transformation by v-src. Mol Cell Biol. 1998;18(5):2553–2558. - PMC - PubMed
    1. Bromberg JF. Wrzeszczynska MH. Devgan G. Zhao Y. Pestell RG. Albanese C. Darnell JE., Jr Stat3 as an oncogene. Cell. 1999;98(3):295–303. - PubMed
    1. Buettner R. Mora LB. Jove R. Activated STAT signaling in human tumors provides novel molecular targets for therapeutic intervention. Clin Cancer Res. 2002;8(4):945–954. - PubMed

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