Xi Tao, Xia Huan, Fan Yu-Xiang, Cao Yong-Cheng, Zhang Hong-Liang
Xinjiang Medical University Urumqi 830000,China.
Science of Nuclear Medicine,Cancer Hospital Affiliated to Xinjiang Medical University Urumqi 830011,China.
Zhongguo Zhong Yao Za Zhi. 2019 Jan;44(1):119-124. doi: 10.19540/j.cnki.cjcmm.20180914.001.
To explore the mechanism of β-carboline alkaloids inhibiting the migration and invasion of SGC-7901 cells and its correlation with FAK gene expression,CCK-8 method was used to determine the inhibitory rate of β-carboline alkaloids on the proliferation of gastric cancer SGC-7901 cells under different concentrations.The effect of β-carboline alkaloids on the migration and invasion of SGC-7901 cells was used by Transwell compartment.Detection of mRNA and protein expression of FAK genes were used by qRT-PCR and Western blot.Then si-FAK-1051 recombinant plasmid was transfected into SGC-7901 cells.FAK gene silencing effect was identified by qRT-PCR and Western blot technique again.Finally,the effects of FAK gene silencing on proliferation and migration of gastric cancer SGC-7901 cells were detected by CCK-8 kit and Transwell chamber assay respectively.With the increase of the concentration ofβ-carboline alkaloids,the inhibitory rate of SGC-7901 cells in human gastric cancer cells increased gradually,with IC5013.364 mg·L-1.The number of SGC-7901 cells of Transwell compartment in the positive experimental group(5-FU,5 mg·L-1) and the β-carboline alkaloids group decreased significantly(P<0.01) and the number of SGC-7901 cells in the β-carboline alkaloids group was significantly lower than that in the positive experimental group(P<0.01).Compared with the blank control group,the mRNA and protein expression level of FAK genes in the positive experimental group was significantly lower than that in the experimental group of β-carboline alkaloids(P<0.05).After transfection of si-FAK-1051 into gastric cancer SGC-7901 cells,the expression of mRNA and protein of FAK gene was significantly down regulated(P<0.05).SGC-7901 cell proliferation and cell migration ability also decreased significantly(P<0.05).β-carboline alkaloids are more effective than 5-FU in inhibiting migration and invasion of gastric cancer SGC-7901 cells,and the mechanism may be related to the inhibition of mRNA and protein expression of FAK gene by β-carboline alkaloids.
为探讨β-咔啉生物碱抑制SGC-7901细胞迁移和侵袭的机制及其与FAK基因表达的相关性，采用CCK-8法检测不同浓度β-咔啉生物碱对胃癌SGC-7901细胞增殖的抑制率。利用Transwell小室检测β-咔啉生物碱对SGC-7901细胞迁移和侵袭的影响。采用qRT-PCR和Western blot检测FAK基因的mRNA和蛋白表达。然后将si-FAK-1051重组质粒转染至SGC-7901细胞中，再次通过qRT-PCR和Western blot技术鉴定FAK基因沉默效果。最后，分别用CCK-8试剂盒和Transwell小室检测FAK基因沉默对胃癌SGC-7901细胞增殖和迁移的影响。随着β-咔啉生物碱浓度的增加，人胃癌SGC-7901细胞的抑制率逐渐升高，IC50为13.364 mg·L-1。阳性实验组（5-FU，5 mg·L-1）和β-咔啉生物碱组Transwell小室中SGC-7901细胞数量均显著减少（P<0.01），且β-咔啉生物碱组SGC-7901细胞数量显著低于阳性实验组（P<0.01）。与空白对照组相比，阳性实验组FAK基因的mRNA和蛋白表达水平显著低于β-咔啉生物碱实验组（P<0.05）。将si-FAK-1051转染至胃癌SGC-7901细胞后，FAK基因的mRNA和蛋白表达显著下调（P<0.05）。SGC-7901细胞增殖和细胞迁移能力也显著降低（P<0.05）。β-咔啉生物碱在抑制胃癌SGC-7901细胞迁移和侵袭方面比5-FU更有效，其机制可能与β-咔啉生物碱抑制FAK基因的mRNA和蛋白表达有关。
