论文部分内容阅读
目的探讨鞘氨醇激酶1(SphKl)通路在食管癌侵袭和转移中的作用及临床意义。方法采用Real-timePCR和Western blot检测SphKl在食管癌组织中的表达,设计构建SphKl靶向shRNA质粒,建立SphKl稳定沉默的细胞系。MTT和Transwell法检测SphKl基因沉默对EC-1细胞增殖、侵袭的影响,明胶酶谱检测其对基质金属蛋白酶-9(MMP-9)和MMP-2分泌的影响。结果 SphKl的mRNA和蛋白质表达水平与侵袭能力明显相关。SphKl-siRNA能显著抑制EC-1细胞的增殖和侵袭,并能显著抑制EC-1细胞的MMP-9和MMP-2蛋白分泌。结论SphKl与食管癌侵袭和转移关系密切,转染靶向SphKl基因的siRNA序列能够抑制食管癌EC-1细胞的增殖和侵袭,其机制与抑制MMP-9和MMP-2蛋白分泌密切相关。
Objective To investigate the role of SphKl pathway in the invasion and metastasis of esophageal cancer and its clinical significance. Methods Real-time PCR and Western blot were used to detect the expression of SphKl in esophageal cancer tissue. The SphKl targeted shRNA plasmid was designed to establish a stable silencing cell line of SphKl. MTT and Transwell assay were used to detect the effect of SphKl gene silencing on the proliferation and invasion of EC-1 cells. Gelatin zymogram was used to detect the effect of MMP-2 and MMP-2 secretion. Results The mRNA and protein expression levels of SphKl were significantly related to the invasive ability. SphKl-siRNA can significantly inhibit the proliferation and invasion of EC-1 cells, and can significantly inhibit the secretion of MMP-9 and MMP-2 proteins from EC-1 cells. Conclusion SphKl is closely related to the invasion and metastasis of esophageal cancer. Transfection of siRNA targeting SphKl gene can inhibit the proliferation and invasion of esophageal cancer EC-1 cells. The mechanism is closely related to the inhibition of the secretion of MMP-9 and MMP-2.