IJMCM (International Journal of Molecular and Cellular Medicine) Summer 2016, Vol 5, No 3.

Genistein Induces Apoptosis and Inhibits Proliferation of HT29 Colon Cancer Cells

Gholamreza Shafiee, et al

Department of Biochemistry, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran; Department of Molecular Medicine and Human Genetics, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

Soybean isoflavone genistein has multiple anticancer properties and its pro-apoptotic and anti-proliferative effects have been studied in different cancer cells. However, the mechanisms of action of genistein and its molecular targets on human colon cells have not been fully elucidated. Therefore, caspase-3 and p38 mitogenactivated protein kinase (p38 MAPK) as the main therapeutic targets were investigated in this study at both gene expression and protein levels in HT29 colon cancer cells. The caspase-3 and p38 MAPK gene expression levels were examined by real time PCR whereas flow cytometry technique was performed to determine their intracellular protein levels. The caspase-3 enzyme activity was obtained by colorimetric method while the gelatinase activity of matrix metalloproteinase-2 (MMP2) was determined by zymography. In addition, MTT test, wound healing assay and clonogenic assay were carried out to determine the effect of genistein on HT29 cell viability, migration, and proliferation, respectively. Genistein induced apoptotic death in HT29 cells through activation of caspase-3 pathway at the transcriptional, protein, and enzymatic levels. Moreover, genistein inhibited the proliferation of HT29 cells by reducing of both p38 MAPK gene expression and its active phosphorylated protein level. Also, we showed that genistein strongly suppressed the metastatic potency of HT29 colon cancer cells via the reduction of MMP2 activity. Based on the results of this study, we conclude that genistein may exhibit its anticancer properties on HT29 colon cancer cells by modulating caspase-3 and p38 MAPK pathway at different transcriptional and protein levels.

伊朗   IJMCM(国际分子和细胞医学杂志)

Summer 2016, Vol 5, No 3.

染料木黄酮诱导细胞凋亡和抑制HT29结肠癌细胞的增殖

Gholamreza Shafiee, et al

哈马丹医学科学院生物化学系，哈马丹，伊朗; 哈马丹医学科学院分子医学和人类遗传学系，哈马丹，伊朗

大豆异黄酮染料木黄酮具有多种抗癌性质，并且已经在不同的癌细胞中研究了其促凋亡和抗增殖作用。然而，染料木黄酮及其分子靶物对人结肠细胞的作用机制尚未完全阐明。因此，本研究在HT29结肠癌细胞的基因表达和蛋白水平上研究caspase-3和p38丝裂原活化蛋白激酶（p38 MAPK）作为主要治疗靶标。通过实时PCR检查半胱天冬酶-3和p38 MAPK基因表达水平，而进行流式细胞术技术以确定其细胞内蛋白质水平。通过比色法获得半胱天蛋白酶-3酶活性，通过酶谱法测定基质金属蛋白酶-2（MMP2）的明胶酶活性。此外，进行MTT测试，伤口愈合测定和克隆形成测定以分别测定染料木黄酮对HT29细胞存活力，迁移和增殖的影响。染料木黄酮通过活化半胱天冬蛋白酶-3途径在转录，蛋白质和酶水平诱导HT29细胞中的凋亡死亡。此外，染料木素通过减少p38 MAPK基因表达和其活性磷酸化蛋白水平抑制HT29细胞的增殖。此外，我们表明染料木黄酮通过减少MMP2活性强烈抑制HT29结肠癌细胞的转移作用。基于该研究的结果，我们得出结论，染料木黄酮可以通过调节半胱天蛋白酶-3和p38 MAPK通路在不同的转录和蛋白水平上显示其对HT29结肠癌细胞的抗癌特性。