Major Project

Increased expression of RRM2 by human papillomavirus E7 oncoprotein promotes angiogenesis in cervical cancer

Wang et al. (2014) conducted a study to investigate roles of ribonucleotide reductase M2 (RRM2) as a novel potential target activated by human papillomavirus (HPV) E7 oncoprotein on cervical cancer progression. In previous studies (Rosty et al., 2005; Kuner et al., 2007) found many novel potential downstream genes for HPVE7 oncoprotein by microarray technique, RRM2 is the one of novel potential downstream genes for HPVE7 oncoprotein. In addition, overexpression RRM2 is also associated with tumor angiogenesis and tumor metastasis (Zhang et al., 2009; Zhou et al., 1998.) The study was divided into three parts; cell culture, animal, and human specimen experiments. In cell culture, HeLa cell line, a positive HPV type 18 cervical cell line which has HPVE7 oncoprotein, and C33A cell line, a negative HPV which has no HPV oncoprotein, were used in the study. C33A cell line was transfected with plasmid for HPVE7 for biochemical characterization as HeLa cell line to confirm results of roles of RRM2 on cervical cancer progression. In animal experiment, BALB/c nude mice were inoculated subcutaneously with 5x10⁶ C33A-transfected RRM2 (C33A-RRM2) and C33A-transfected empty vector (C33A-V). In human specimen experiment, human cervical cancer samples were collected from cervical cancer patients to stain immunohistochemistry for RRM2 and CD31, an angiogenesis biomarker, expressions. The results showed RRM2 as a novel target for HPVE7 to mediated cervical cancer progression in all parts of experiment.  Therefore, RRM2 may be a therapeutic target for cervical cancer treatment or a biomarker for predicting the prognosis of cervical cancer in the future.

       This provides a novel target downstream activated of HPVE7 to mediate cervical cancer progression. However, there are some limitations.

1.  This study chose only positive HPV type 18 cancer cell line (HeLa cells). The research may choose other positive HPV type 16 cancer cell lines such as CaSki cells to confirm their results.

2.  For animal experiment, the study used nude mice-transplanted C33A-RRM2 only to determine the roles of RRM2 in cervical cancer progression. The study may use the posivtive HPVE7 cell line together to confirm the in vivo results.

3.   For human specimen, the study should determine correlation between RRM2 expression and cervical cancer stage for more information for predicting the prognosis of cervical cancer in the future.

The strength of the study is that the confirmation of supporting roles of RRM2 in cervical cancer progression in positive HPVE7 HeLa cell line by comparing to negative HPVE7 C33A cell line which were transfected with RRM2.

References:

     1. Wang, N., Zhan, T., Ke, T., Huang, X., Ke, D., Wang, Q., & H, Li. (2014). Increased expression of RRM2 by human papillomavirus E7 oncoprotein promotes angiogenesis in cervical cancer. Br J Cancer. 18; 110(4):1034-44.

     2. Rosty, C., Sheffer, M., Tsafrir, D., Stransky, N., Tsafrir, I., Peter, M., & et al. (2005). Identification of a proliferation gene cluster associated with HPV E6/E7 expression level and viral DNA load in invasive cervical carcinoma. Oncogene. 27; 24(47):7094-104.

   3. Kuner, R., Vogt, M., Sultmann, H., Buness, A., Dymalla, S., Bulkescher, J., & et al. (2007). Identification of cellular targets for the human papillomavirus E6 and E7 oncogenes by RNA interference and transcriptome analyses. J Mol Med (Berl). 85(11):1253-62. Epub 2007 Jun 23.

    4. Zhang, K., Hu, S., Wu, J., Chen, L., Lu, J., Wang, X., et al. (2009). Overexpression of RRM2 decreases thrombspondin-1 and increases VEGF production in human cancer cells in vitro and in vivo: implication of RRM2 in angiogenesis. Mol Cancer. 28;8:11.

     5. Zhou, B.,S., Tsai, P., Ker, R., Tsai, .J, Ho, R’, Yu, J., et al. (1998). Overexpression of transfected human ribonucleotide reductase M2 subunit in human cancer cells enhances their invasive potential. Clin Exp Metastasis. 16(1):43-9.

Minor Assignment

           My research question is the following; what are the impacts of mesenchymal stem cells (MSCs) on tumor proliferation and tumor angiogenesis in cervical cancer cells-transplanted into nude mice. Nowadays, a number of clinical trials on MSCs therapy have been rising since 2004, especially tissue repair because MSCs can possess self-renewal and can be differentiated into ectoderm, mesoderm, and endoderm and also can home to inflammatory sites such as wound. Tumor environment is characterized by the presence of inflammatory cells as well as wound. However, the impact of MSCs on tumor environment remains controversial.

           Researchers who have looked at this subject are Huang and co-worker, and Kéramidas and co-worker. Both of studies focused on the impact of MSCs on tumor growth and tumor angiogenesis, although they did experiments in the different cancer models. These results were interesting evidences for research in this filed as well.

         Huang and co-worker (2013) argue that MSCs increased the tumor growth and tumor angiogenesis through interleukin-6/endothelin-1/alternatively protein B (IL-6/ET-1/Akt) pathway by activating MSCs-secreted IL-6 in human colorectal cancer cells-transplanted nude mice.

     In contrast, Kéramidas and co-worker (2013) investigate the impact of MSCs in adenocarcinoma cells-transplanted nude mice model. They found MSCs decreased tumor growth. Although they could not decrease tumor angiogenesis, they could normalize tumor vasculature.

         The results of both studies were controversial. Therefore, my study aims to clarify this issue in cervical cancer cell-transplanted nude mice model.

        My work will be closer to Huang’s and Kéramidas’ because both studies demonstrated the impact of MSCs on colorectal cancer and adenocarcinoma cancer, respectively. My study will investigate the impact of MSCs on tumor growth, tumor angiogenesis, and involved mechanisms in cervical cancer-transplanted nude mice model.

       The findings of my study may be a new knowledge for further experiments or cancer therapies.

References:

1. Huang, W.H., Chang, M.C., Tsa, K.S., Hung, M.C., Chen, H.L., & Hung, S.C. (2013). Mesenchymal stem cells promote growth and angiogenesis of tumors in mice. Oncogene, 12;32(37):4343-54.

2. Kéramidas, M., Fraipont, F., Karageorgis, A., Moisan, A., Persoons, V., Richard, M., & et al. (2013). The dual effect of mesenchymal stem cells on tumour growth and tumour angiogenesis. Stem Cell Research & Therapy, 4:41. 

Assignment2 (Introduction)

The anti-tumor effect of curcumin on pRb degradation and plasma protein extravasation releasing in cervical cancer-implanted nude mice model

Natchaya Wongeakin

Stage1: Almost all cases of cervical cancer which are attributed by HPV infection is the fourth most common cancer in women worldwide. It is also the fourth most common cause of cancer death (266,000 deaths in 2012); almost 70% of cases are in areas with lower levels of development (WHO, 2013). Nowadays, ICO HPV Information Centre estimates that every year 8,184 Thai women are diagnosed with cervical cancer and 4,513 Thai women die from the disease. Cervical cancer ranks as the 2nd most common cancer in Thai women, especially women between 15 and 44 years of age (ICO Information Centre on HPV and Cancer, 2014).

Satge2: The major oncogenic HPV protein E7 is involved in the immortalization of target cells by inactivation of cellular tumor suppressor retinoblastoma protein (pRb), leading to the release of activation of E2f (Huh et al, 2007). The pRb degradation can enhance angiogenic factors through HIF pathway leading to cancer cell proliferation and differentiation, and cancer nrovascularization (Duxbury and Whang, 2007; Wang et al 2014). In addition to angiogenic factors, HIF pathway is also associated with endothelial progenitor cell (EPC) migration to mediate cancer neovascularization (Matsui et al, 2004; Litz et al, 2006) by motivating of plasma protein extravasation releasing such as SDF-1 (Ceradini et al, 2004; kioi et al, 2010). In recently, plants contain numerous bioactive molecules such as polyphenols are used for prevention and treatment in various diseases. Curcumin is the one of polyphenols which several researches indicated the anti-tumorigenic property (Basu et al, 2013; Dai et al, 2013; Yin et al, 2013).
 

Stage3: However, it is unclear whether curcumin could decrease cervical cancer progression by inhibiting pRb degradation and plasma protein extravasation releasing in cervical cancer.

Stage4: In the present study aim to investigate the anti-tumor effects of curcumin on pRb and SDF-1 expressions in cervical cancer-implanted nude mice model.

Stage5: These findings would identify a novel treatment to decrease cervical cancer progression.

Assignment1: Citation

Matrix metalloproteinase-10 promotes tumor progression through regulation of angiogenic and apoptotic pathways in cervical tumors

Abstract

Background: Cancer invasion and metastasis develops through a series of steps that involve the loss of cell to cell and cell to matrix adhesion, degradation of extracellular matrix and induction of angiogenesis. Different protease systems (e.g., matrix metalloproteinases, MMPs) are involved in these steps. MMP-10, one of the lesser studied MMPs, is limited to epithelial cells and can facilitate tumor cell invasion by targeting collagen, elastin and laminin. Enhanced MMP-10 expression has been linked to poor clinical prognosis in some cancers, however, mechanisms underlying a role for MMP-10 in tumorigenesis and progression remain largely unknown. Here, we report that MMP-10 expression is positively correlated with the invasiveness of human cervical and bladder cancers.

Methods: Using commercial tissue microarray (TMA) of cervical and bladder tissues, MMP-10 immunohistochemical staining was performed. Furthermore using a panel of human cells (HeLa and UROtsa), in vitro and in vivo experiments were performed in which MMP-10 was overexpressed or silenced and we noted phenotypic and genotypic changes.

Results: Experimentally, we showed that MMP-10 can regulate tumor cell migration and invasion, and endothelial cell tube formation, and that MMP-10 effects are associated with a resistance to apoptosis. Further investigation revealed that increasing MMP-10 expression stimulates the expression of HIF-1α and MMP-2 (pro-angiogenic factors) and PAI-1 and CXCR2 (pro-metastatic factors), and accordingly, targeting MMP-10 with siRNA in vivo resulted in diminution of xenograft tumor growth with a concomitant reduction of angiogenesis and a stimulation of apoptosis.

Conclusion: Taken together, our findings show that MMP-10 can play a significant role in tumor growth and progression, and that MMP-10 perturbation may represent a rational strategy for cancer treatment.

Keywords: Angiogenesis, Apoptosis, Cancer, Invasion, MMP-10

Reference

Zhang G, Miyake M, Lawton A, Goodison S,  Rosser CJ. Matrix metalloproteinase-10 promotes tumor progression through regulation of angiogenic and apoptotic pathways in cervical tumors. BMC Cancer. 2014, 14:310. 

Results and findings

MMP-10 is the important factor to promote tumor growth and progression through enhanced tumor cell migration and invasion, angiogenesis, and apoptosis resistance. Interestingly, MMP-10 may be a new molecular target for cancer therapy.

Citations

1. Zhang et al. (2014) reported that both MMP-10 activities in vivo and in vitro experiments were shown to be important role in the tumor growth through upregulated key molecules related to angiogenesis, metastasis, and apoptosis resistance.

2. Zhang et al. (2014) revealed that MMP-10 with siRNA could decrease tumor growth in vivo experiment that is associated with a reduction of angiogenesis and a stimulation of apoptosis.

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