Jahanban-Esfahlan Rana, Seidi Khaled, Monfaredan Amir, Shafie-Irannejad Vahid, Abbasi Mehran Mesgari, Karimian Ansar, Yousefi Bahman
Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran; Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran; Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Gene. 2017 May 20;613:14-19. doi: 10.1016/j.gene.2017.02.034. Epub 2017 Mar 1.
Earlier, we verified that Melissa officinalis extract (MOE) elicits potent antiproliferative effects on different human cancer cells. To gain insights into the molecular mechanisms accounting for the cytotoxic effects of MOE, we assessed the expression patterns of several prominent molecules with therapeutic potential in cancer by Quantitative PCR (Q-PCR).
A549, MCF-7 and PC3 cancer cells were grown in complete RPMI 1640 and seeded in 24 well micro plates. After incubation for 72h, 100μg/ml of MOE was added and the cells were further incubated for 72h. Afterwards, the cells were subjected to RNA extraction for the means of Q-PCR.
Our results indicated that in PC3 cancer cells, MOE resulted in a significant downregulation of VEGF-A (0.0004 fold), Bcl-2 (0.001 fold), Her2 (0.02 fold), and hTERT (0.023 fold) compared to the untreated control. In addition, VEGF-A and hTERT mRNA were significantly downregulated in MCF-7 and A549 cancer cells, as well. Notably, high anti-angiogenic activity was closely associated with a high anti-telomerase activity of MOE in studying cancer cells. The decrease in VEGF-A expression was significantly superior than that of hTERT downregulation, as PC3 cancer cells with the highest hTERT down regulation (0.023) presented the highest anti VEGF activity (0.0004 fold), whereas MCF-7 cells with the lowest hTERT inhibition (0.213) showed the lowest VEGF inhibition(0.0435) among the three studied cancer cells. We noticed that the modulation of VEGF-A and hTERT gene expression can be considered as a common target, accounting for the therapeutic potential of MOE on human breast, lung and prostate cancer cells.
Altogether, it is suggested that the potent antiproliferative activity of the hydroalcoholic extract of Melissa officinalis is somehow explainable by its high potency to inhibit expression of the prominent oncogenes Bcl2, Her2, VEGF-A and hTERT in prostate cancer. In tumors with functional p53, including MCF-7 and A549 cancer cells, the role of p53, Bcl2 and Her2 is less significant. It appears that MOE exerts its antiproliferative effects in these cancer cells partly via concurrent downregulation of VEGF-A and hTERT. Additional studies are needed to clarify the role of other active molecules in cancer cells harboring functional p53.
此前，我们证实了蜜蜂花提取物（MOE）对不同的人类癌细胞具有强大的抗增殖作用。为深入了解MOE细胞毒性作用的分子机制，我们通过定量PCR（Q-PCR）评估了几种在癌症治疗中具有潜在作用的重要分子的表达模式。
A549、MCF-7和PC3癌细胞在完全RPMI 1640培养基中培养，并接种于24孔微孔板中。孵育72小时后，加入100μg/ml的MOE，细胞再孵育72小时。之后，对细胞进行RNA提取用于Q-PCR分析。
我们的结果表明，在PC3癌细胞中，与未处理的对照相比，MOE导致VEGF-A（0.0004倍）、Bcl-2（0.001倍）、Her2（0.02倍）和hTERT（0.023倍）显著下调。此外，VEGF-A和hTERT mRNA在MCF-7和A549癌细胞中也显著下调。值得注意的是，在研究癌细胞时，MOE的高抗血管生成活性与高抗端粒酶活性密切相关。VEGF-A表达的降低明显优于hTERT下调，因为hTERT下调最高（0.023）的PC3癌细胞呈现出最高的抗VEGF活性（0.0004倍），而在三种研究的癌细胞中，hTERT抑制最低（0.213）的MCF-7细胞显示出最低的VEGF抑制（0.0435）。我们注意到，VEGF-A和hTERT基因表达的调节可被视为一个共同靶点，这解释了MOE对人乳腺癌、肺癌和前列腺癌细胞的治疗潜力。
总之，提示蜜蜂花水醇提取物强大的抗增殖活性在某种程度上可归因于其高效抑制前列腺癌中重要癌基因Bcl2、Her2、VEGF-A和hTERT表达的能力。在具有功能性p53的肿瘤中，包括MCF-7和A549癌细胞，p53、Bcl2和Her2的作用不太显著。似乎MOE在这些癌细胞中部分通过同时下调VEGF-A和hTERT发挥其抗增殖作用。需要进一步研究以阐明其他活性分子在具有功能性p53的癌细胞中的作用。
