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. 2013 Jun;5(6):1777-1782.
doi: 10.3892/ol.2013.1295. Epub 2013 Apr 8.

Treatment with Tie2-siRNA in combination with carboplatin suppresses the growth of Ishikawa human endometrial carcinoma cell xenografts in vivo

Feifei Guo  1 , Qingying Xun, Huaijun Zhou
Affiliations

Treatment with Tie2-siRNA in combination with carboplatin suppresses the growth of Ishikawa human endometrial carcinoma cell xenografts in vivo

Feifei Guo et al. Oncol Lett. 2013 Jun.

Abstract

It is well-known that tumor angiogenesis is important in cancer development, and studies on blocking angiogenesis to treat tumors have become one of the most promising and active fields in anticancer research. The present study investigated the effect of siRNA targeting the tyrosine kinase receptor 2 (Tie2) gene in combination with carboplatin in a mouse model of endometrial carcinoma in an attempt to elucidate the role of Tie2 in the carcinogenesis and progression of endometrial carcinoma via angiogenesis, in order to establish a basis for the development of complementary molecule targeting and chemotherapeutic actions. Ishikawa cells were used to establish a human endometrial carcinoma nude mouse tumor xenograft model. Tie2-siRNA (20 μg/mouse) and/or carboplatin (25.0 mg·kg-1) were administered as the treatment strategy. Real-time PCR and western blotting were used to evaluate the expression levels of Tie2 mRNA and protein and immunohistochemistry was used to assess the vessel density of the tumor tissues. The present data demonstrated that Tie2-siRNA and/or carboplatin were able to suppress the growth of endometrial xenografts in vivo and attenuate the expression of Tie2 mRNA and protein, as assessed by real-time PCR and western blotting. Furthermore, immunohistochemical assessment showed that the vessel density of the tumors decreased with treatment. The present results suggest that treatment with Tie2-siRNA or carboplatin alone was able to inhibit the growth of human endometrial carcinoma nude mouse xenografts markedly and decrease the expression of Tie2. The combination of Tie2-siRNA and carboplatin increased the therapeutic effect of carboplatin which may eliminate the tumor microenvironment, increase the apoptosis of tumor cells, normalize the abnormal tumor vessels and increase the efficiency of chemotherapy for endometrial carcinoma with carboplatin. The synergy of Tie2-siRNA in combination with carboplatin may involve the regulation of other angiogenesis and metastasis pathways.

Keywords: carboplatin; endometrial carcinoma; nude mice; siRNA; tyrosine kinase receptor 2.

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Figures

Figure 1
Figure 1
Tie2-siRNA and/or carboplatin affects tumor growth in vivo. (A) Macroscopic appearance of tumors in nude mice 30 days after the transplantation of human endometrial carcinoma Ishikawa cells. Arrows outline the representative tumors in the right scapular region of the mice. (B) Images of representative excised tumors from each group. (C) Tumor volumes were averaged for each treatment group and time point over the course of the study. (D) Final tumor volumes of the isolated tumors were averaged for each group (mean ± SEM). The tumor volumes of mice treated with of carboplatin, Tie2-siRNA and the combined administration were significantly different, compared with the G or N group, beginning at the third treatment and continuing to be significant throughout the study (P<0.05 respectively). aCompared with G group, there was a statistically significant difference; bCompared with N group, there was a statistically significant difference; cCompared with C group, there was a statistically significant difference; dCompared with T group, there was a statistically significant difference. Tie2, tyrosine kinase receptor 2; G, glucose treatment; N, pRNAT-CMV3.2-Neo carrying negative siRNA treatment; C, carboplatin treatment; T, pRNAT-CMV3.2-Neo carrying Tie2-siRNA treatment; A, Tie2-siRNA in combination with carboplatin treatment.
Figure 2
Figure 2
Tie2-siRNA and/or carboplatin decreases Tie2 expression in the tumor tissues of the groups. (A) Total RNA was isolated from tumor tissues and relative Tie2 mRNA expression was analyzed with real-time PCR. (B) Western blot analysis of Tie2 protein expression in five groups and (C) quantification of relative Tie2 protein expression tumor tissues of every group (P<0.05). aCompared with G group, there was a statistically significant difference; bCompared with N group, there was a statistically significant difference; cCompared with C group, there was a statistically significant difference; dCompared with T group, there was a statistically significant difference. Tie2, tyrosine kinase receptor 2; G, glucose treatment; N, pRNAT-CMV3.2-Neo carrying negative siRNA treatment; C, carboplatin treatment; T, pRNAT-CMV3.2-Neo carrying Tie2-siRNA treatment; A, Tie2-siRNA in combination with carboplatin treatment.
Figure 3
Figure 3
Inhibition of tumor angiogenesis following treatment. (A) Immunohistochemical staining of tumor xenografts probed with anti-CD34 antibody after adminastration to visualize endothelial cells and tumor vascular formation. The images of the microvessel density and vessel characteristics of the treatment groups are shown (magnification, ×200). (B) Results of immunohistochemical staining. The densitiy of tumor vessels were lower in the carboplatin, Tie2-siRNA and the combined administration groups than those in the G and N groups (P<0.05). The combined administration group showed the greatest reduction in the number of tumor vessels and the reduction was significantly different compared with the other groups (P<0.05). aCompared with G group, there was a statistically significant difference; bCompared with N group, there was a statistically significant difference; cCompared with C group, there was a statistically significant difference; dCompared with T group, there was a statistically significant difference. Tie2, tyrosine kinase receptor 2; G, glucose treatment; N, pRNAT-CMV3.2-Neo carrying negative siRNA treatment; C, carboplatin treatment; T, pRNAT-CMV3.2-Neo carrying Tie2-siRNA treatment; A, Tie2-siRNA in combination with carboplatin treatment.
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References

    1. National Cancer Institute at the National Institutes of Health Endometrial Cancer Home Page. http://seer.cancer.gov/statfacts/html/corp.html Accessed November 10, 2010.
    1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90. - PubMed
    1. Myatt SS, Wang J, Monteiro LJ, Christian M, Ho KK, Fusi L, Dina RE, Brosens JJ, Ghaem-Maghami S, Lam EW. Definition of microRNAs that repress expression of the tumor suppressor gene FOXO1 in endometrial cancer. Cancer Res. 2010;70:367–377. - PMC - PubMed
    1. Ray M, Fleming G. Management of advanced-stage and recurrent endometrial cancer. Semin Oncol. 2009;36:145–154. - PubMed
    1. Poon RT, Lau CP, Ho JW, Yu WC, Fan ST, Wong J. Tissue factor expression correlates with tumor angiogenesis and invasiveness in human hepatocellular carcinoma. Clin Cancer Res. 2003;9:5339–5345. - PubMed

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