新型铂配合物Mor-platin导致细胞凋亡及抑制细胞迁移

Mor-platin a new platinum complex that can induce apoptosis and inhibit migration

探讨了新型铂配合物Mor-platin对人肺癌细胞(H1975)的抗癌效果以及对细胞迁移和侵袭的影响.运用噻唑蓝法和活性氧检测试剂盒,比较Mor-platin与经典抗癌药物顺铂影响H1975细胞存活率和活性氧产生的不同;利用细胞周期与细胞凋亡检测试剂盒,研究Mor-platin和顺铂对细胞凋亡与细胞周期的影响;Western blot法检测Mor-platin对caspase相关蛋白caspase 3/8/9/脱氧核糖核酸修复酶(PARP)蛋白水平和凋亡相关蛋白B-淋巴瘤-2基因相关蛋白(Bax)/B-淋巴瘤-2基因(Bcl-2)比值的改变;最后使用划痕实验和transwell装置研究Mor-platin和顺铂对H1975细胞迁移和侵袭的影响.实验结果显示,Mor-platin与顺铂相比,在更低的浓度下即能杀死H1975细胞.另外,Mor-platin比顺铂更易产生细胞内活性氧,使细胞阻滞于S期,激活caspase相关蛋白caspase 3/8/9/PARP,增加Bax/Bcl-2比值,最终导致细胞凋亡.通过划痕与transwell实验还发现,Mor-platin可以抑制肿瘤细胞迁移和侵袭,而顺铂没有这种现象.综上所述,Mor-platin具有更好的抗肺癌细胞能力,以及独特的抑制细胞迁移能力,为铂类化合物的开发和肿瘤药物的研究提供了新思路.

Since cisplatin was discovered and applied clinically, people have synthesized thousands of platinum compound derivatives. These platinum derivatives are effective in chemotherapy for some tumors including testicular cancer, ovarian cancer, lung cancer, bladder cancer, head and neck cancer. All of these platinum complex are believed to exert their unparalleled anticancer potency by interacting with nulear DNA, resulting in DNA adducts formation and consequently lead to cell apoptosis, cell cycle arrest and reactive oxygen species released. However, these platinum complex are severely limited by drug resistance and undesirable side effects arising from relatively low specificity and resultant high dosage requirement. To overcome the drug-resistance and improve therapeutic efficiency, it is necessary to synthesized new platinum complex. Our group successfully synthesized a new platinum complex Mor-platin, it can significantly damage more tumor cells than traditional drug cisplatin, while the other biological effects have not been studied. This work aims to study the effect of Mor-platin on its anti-cancer ability, cell migration and cell invasion. In order to-detect anti-cancer efficiency, MTT assay was used, and the results showed that IC150 of Mor-platin（14.41 mg mL^-1） was-far less than IC50 of cisplatin（97.29mg mL^-1）, suggesting that Mor-platin have potentially higher tumor cytotoxicity than cisplatin. Next, we used reactive oxygen species assay kit, western blot analysis, apoptosis and cell cycle kit to assess whether Mor-platin can cause cell death by apoptosis, cell cycle, ROS, apoptosis-related protein and caspase protein. The results showed that Mor-platin can increase level of ROS, arrest cell cyle in S phase, increase ratio of Bax/Bcl-2 and activate caspase 3/8/9 resulting in cleaved PARP, ultimately lead to cell death. The major threat of cancer is not from the primary tumor itself but from metastasis to other organs. To develop successful metastases, cancer cells need to disseminate to a distant organ, migrate out of the primary tumor, invade to blood or lymphatic vessels, finally seed, proliferate and colonize to form secondary tumors. Cell migration is a basic process within the cancer. Tumor metastasis is a major cause of cancer deaths, 90% of the cancer deaths can be attributed to the tumor metastasis, therefore it is necessary to avoid tumor metastasis on clinical cancer treatment. Only understanding the clear tumor metastasis for emerging antitumor drugs, they can be obtained to development and application. Current tumor metastasis experiments include transwell assay, wound-healing assay, microcarrier beads, three-dimensional assay and so on. Here, we used wound-healing assay and transwell assay to quantify the migration inhibitory activity of Mor-platin. Intriguingly, both wound-healing assay and transwell assay showed that Mor-platin can also inhibit cell migration and invasion, while cisplatin did not show any same effects. In summary, this new platinum complex Mor-platin not only have high anti-cancer efficiency but also inhibit migration, providing a novel strategy for the development of next generation platinum complex with optimal anti-tumor activities.