木棉皮抗消化道肿瘤活性部位筛选及抑制肿瘤转移机制

Screening of the Active Fraction of Bombax ceiba against Gastrointestinal Tumors and Its Mechanism of Inhibiting Tumor Metastasis

为筛选木棉皮醇提物抗消化道肿瘤活性部位,探究其抑制敏感肿瘤细胞增殖、转移作用机制,通过采用CCK-8(cell counting kit-8)法考查木棉皮醇提物不同极性萃取部位对人肝癌HepG2细胞、人胃癌SGC7901细胞、人结肠癌SW480细胞和人胰腺癌PANC-1细胞这4种肿瘤细胞的抑制作用,筛选出木棉皮醇提物抗肿瘤的活性部位和敏感细胞。采用细胞划痕实验、Transwell实验和细胞黏附实验,研究木棉皮醇提物抗肿瘤活性部位对敏感肿瘤细胞迁移、侵袭和黏附能力的影响。定量聚合酶链反应法(quantitative polymerase chain reaction,qPCR)检测基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)基因的mRNA转录水平。结果表明,木棉皮醇提物不同极性萃取部位中石油醚部位对人胃癌SGC7901细胞和人肝癌HepG2细胞的抑制率最大,且对人胃癌SGC7901细胞最为敏感。随着木棉皮醇提物石油醚部位浓度增加,在24、48、72 h时间段人胃癌SGC7901细胞的迁移面积相对于空白组有所减小(P<0.05)。与空白组相比,木棉皮醇提物石油醚萃取部位有抑制肿瘤细胞侵袭能力且在一定浓度范围呈剂量依赖性(P<0.05);随着木棉皮醇提物石油醚萃取部位浓度增加,细胞黏附抑制率逐渐提高(P<0.05)。木棉皮醇提物石油醚萃取部位可明显抑制MMP-2和MMP-9基因的mRNA转录水平(P<0.05)。可见,木棉皮醇提物石油醚萃取部位可通过调控MMP-2和MMP-9基因表达抑制人胃癌SGC7901细胞增殖、迁移、侵袭和黏附,提示该部位可能有抑制转移性胃癌细胞引起的继发性肿瘤作用。

In order to screening the active parts from the ethanol extract of bark of Bombax ceiba against gastrointestinal tumors,to investigate its inhibitory effect on the proliferation and metastasis of sensitive tumor cells.By using cell counting kit-8(CCK-8)examined the inhibitory effects of the ethanol extract of different extraction parts of bark of Bombax ceiba on human hepatocellular carcinoma HepG2 cells,human gastric cancer SGC7901 cells,human colon cancer SW480 cells and human pancreatic cancer PANC-1 cells,the antitumor active parts and sensitive cells of the ethanol extract of bark of Bombax ceiba were screened.Wounding healing assay,transwell migration assay and cell adhesion assay were used to study the effects of anti-tumor active parts of the ethanol extract of bark of Bombax ceiba on migration,invasion and adhesion of sensitive tumor cells.Quantitative polymerase chain reaction(qPCR)method detected mRNA transcription levels of Matrix metalloproteinase-2(MMP-2)and Matrix metalloproteinase-9(MMP-9)genes.The results show that in different polar extraction sites of alcohol extract from Bombax ceiba bark,petroleum ether fraction showed the highest inhibitory rate on human gastric cancer SGC7901 cells and human liver cancer HepG2 cells,and it was most sensitive to human gastric cancer SGC7901 cells.Compared with the blank group,the migration area of human gastric cancer SGC7901 cells decreased at 24 h,48 h and 72 h as the concentration of petroleum ether in the ethanol extract of bark of Bombax ceiba increased(P<0.05).The petroleum ether extract from bark of Bombax ceiba inhibited tumor cell invasion in a dose-dependent manner in a range of concentrations(P<0.05).With the increase of the concentrations of petroleum ether in the ethanol extract of bark of Bombax ceiba,the inhibition rate of cell adhesion increased gradually(P<0.05).The petroleum ether extract from Bombax ceiba bark inhibited mRNA transcription levels of MMP-2 and MMP-9 genes obviously(P<0.05).It is concluded that the petroleum ether extract from bark of Bombax ceiba significantly inhibited the proliferation,migration,invasion and adhesion of human gastric cancer SGC7901 cells by regulating MMP-2 and MMP-9 genes expression,suggested that the extract may inhibit the secondary tumor caused by metastatic gastric cancer cells.