Zhou Wei, Zhou Ling, Wang Min, Liu Zeming, Chen Danyang, Guo Liang
Department of Plastic Surgery, Zhongnan Hospital of Wuhan University ， Wuhan , Hubei , 430071, China.
J BUON. 2020 Jan-Feb;25(1):479-484.
Melanoma is one of the prevalent types of cancer and ranks 6th major cause of cancer associated mortality. In this study the anticancer effects of the carbazole alkaloid Heptazoline were investigated against a panel of melanoma cells.
The normal BJ-5TA and melanoma cell lines MEL-CLS-1M MEL-CLS-2, MEL-CLS-3 were used in this study. MTT and colony formation assays were used to determine the proliferation rate of melanoma cells Aciridine orange (AO)/ ethidium bromide (EB) and annexin V/propidium iodide (PI) staining were used to check the apoptotic cell death. Cell cycle analysis was performed by flow cytometry and protein expression was checked by western blotting.
Heptazoline inhibited the growth of all the melanoma cell lines, exhibiting an IC50 of 15 to 40 µM against the melanoma cells. However, the normal skin cells had IC50 125 µM. The anticancer effects were found to be due to induction of apoptotic cell death which was associated with the upregulation of Bax, cleaved caspase 3, 9 and PARP and downregulation of Bcl-2. Furthermore, Heptazoline also triggered the G0/G1 arrest of melanoma cells. The effects of Heptazoline on the MAPK signalling pathway revealed that this molecule could inhibit the expression of p-p38 concentration-dependently.
Taken together, Heptazoline may prove a lead molecule in the development of systemic therapy of melanoma.
黑色素瘤是常见的癌症类型之一，是癌症相关死亡率的第六大主要原因。在本研究中，对咔唑生物碱庚唑啉针对一组黑色素瘤细胞的抗癌作用进行了研究。
本研究使用了正常的BJ-5TA细胞以及黑色素瘤细胞系MEL-CLS-1M、MEL-CLS-2、MEL-CLS-3。采用MTT和集落形成试验来测定黑色素瘤细胞的增殖率。使用吖啶橙（AO）/溴化乙锭（EB）和膜联蛋白V/碘化丙啶（PI）染色来检查凋亡细胞死亡情况。通过流式细胞术进行细胞周期分析，并通过蛋白质印迹法检测蛋白质表达。
庚唑啉抑制了所有黑色素瘤细胞系的生长，对黑色素瘤细胞的IC50为15至40μM。然而，正常皮肤细胞的IC50为125μM。发现其抗癌作用是由于诱导凋亡细胞死亡，这与Bax、裂解的半胱天冬酶3、9和PARP的上调以及Bcl-2的下调有关。此外，庚唑啉还引发了黑色素瘤细胞的G0/G1期阻滞。庚唑啉对MAPK信号通路的影响表明，该分子可以浓度依赖性地抑制p-p38的表达。
综上所述，庚唑啉可能被证明是黑色素瘤全身治疗开发中的先导分子。
