Xia Chengxing, Luan Ting, Chen Yan, Yan Ruping, Yuan Shunhui, Yang Delin, Wang Haifeng
Department of Urology, The Second Affiliated Hospital of Kunming Medical University, Yunnan Institute of Urology, Kunming 650101, China.
Transl Cancer Res. 2019 Feb;8(1):120-129. doi: 10.21037/tcr.2019.01.12.
Cobra venom membrane toxin (MT) has been defined as a major subset of cobra venom having cardiac toxicity and anticancer activity properties. In our previous study, cobra venom membrane toxin 12 (MT-12), isolated from the snake venom of Chinese , was confirmed to selectively suppress the proliferation and invasion of the bladder cancer (BC) cell line EJ. However, the results have never been confirmed in other bladder cell lines, and the underlying mechanism by which MT-12 inhibits BC is still unknown. Thus, in this study, the effect of MT-12 on the proliferation, adhesion, and invasion of BC cells was explored and . As tumor angiogenesis is a prerequisite for tumor growth and metastasis, the factors involved, such as matrix metalloproteinases (MMPs), vascular endothelial growth factor (VEGF), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1), were tested in our study.
Using RT4 and T24 cells for experiments, CCK-8 assays were used to determine cell proliferation. Annexin V-FITC/PI was used to determine cell apoptosis status. Wound-healing assays were used to determine cell invasion. Cell adhesion experiments were used to determine cell adhesion. Gelatin zymography was used to determine the enzymatic activity of MMP-9 and MMP-2. RT-PCR, ELISA, and immunohistochemistry were used to determine the expression of VEGF, ICAM-1, and VCAM-1.
MT-12 inhibited proliferation, invasion, and adhesion and promoted cell apoptosis in RT4 and T24 cells; this anticancer effect was concentration-dependent. In the BC xenograft mouse model, the results revealed that MT-12 might decrease tumor growth and weight. MT-12 was shown to have an inhibitory effect on MMP-9 activation and the expression of VEGF and ICAM-1 in BC cells and .
The results of the present study, suggest that MT-12 could effectively inhibit BC cell growth and metastasis via inhibition of tumor angiogenesis. As a result, MT-12 may become a novel drug for BC.
眼镜蛇毒膜毒素（MT）已被定义为眼镜蛇毒的一个主要亚群，具有心脏毒性和抗癌活性。在我们之前的研究中，从中华眼镜蛇蛇毒中分离出的眼镜蛇毒膜毒素12（MT-12）被证实可选择性抑制膀胱癌细胞系EJ的增殖和侵袭。然而，这些结果从未在其他膀胱癌细胞系中得到证实，MT-12抑制膀胱癌的潜在机制仍不清楚。因此，在本研究中，探讨了MT-12对膀胱癌细胞增殖、黏附和侵袭的影响。由于肿瘤血管生成是肿瘤生长和转移的先决条件，本研究检测了相关因子，如基质金属蛋白酶（MMPs）、血管内皮生长因子（VEGF）、细胞间黏附分子-1（ICAM-1）和血管细胞黏附分子-1（VCAM-1）。
以RT4和T24细胞为实验对象，采用CCK-8法检测细胞增殖。采用Annexin V-FITC/PI法检测细胞凋亡状态。采用划痕实验检测细胞侵袭。采用细胞黏附实验检测细胞黏附。采用明胶酶谱法检测MMP-9和MMP-2的酶活性。采用RT-PCR、ELISA和免疫组化法检测VEGF、ICAM-1和VCAM-1的表达。
MT-12抑制RT4和T24细胞的增殖、侵袭和黏附，并促进细胞凋亡；这种抗癌作用呈浓度依赖性。在膀胱癌异种移植小鼠模型中，结果显示MT-12可能降低肿瘤生长和重量。MT-12对膀胱癌细胞中MMP-9的激活以及VEGF和ICAM-1的表达具有抑制作用。
本研究结果表明，MT-12可通过抑制肿瘤血管生成有效抑制膀胱癌细胞的生长和转移。因此，MT-12可能成为一种新型的膀胱癌治疗药物。
