纤维粘连蛋白EDA片段调控结肠癌SW480细胞的增殖、凋亡

目的研究纤维粘连蛋白EDA片段对结肠癌SW480细胞增殖、凋亡的影响。方法慢病毒感染结肠癌SW480细胞干扰纤维粘连蛋白EDA片段表达,建立纤维粘连蛋白EDA片段低表达的SW480细胞。利用MTT检测干扰前后细胞增殖的变化,并采用流式细胞术检测干扰前后细胞凋亡的变化。同时使用Western blot分别检测干扰前后凋亡相关蛋白Bax和Bcl-2的表达变化。结果慢病毒感染结肠癌SW480细胞后,纤维粘连蛋白EDA片段持续低表达。MTT法结果显示,干扰纤维粘连蛋白EDA片段24、48、72、96 h后细胞生长速度明显降低。转染组与未转染组及空载体组比较,差异均有统计学意义(P<0.05)。流式细胞术显示未转染组、空载体组及转染组的细胞凋亡率分别为(3.09±1.13)%、(3.71±0.42)%、(47.76±1.51)%,差异有统计学意义(P<0.05)。Western blot的结果显示干扰纤维粘连蛋白EDA片段后促凋亡因子Bax表达上调,同时,凋亡抑制因子Bcl-2表达降低。结论纤维粘连蛋白EDA片段对结肠癌SW480细胞具有促增殖、抗凋亡作用。

Fibronectin EDA domain regulates proliferation and apoptosis of colorectal cancer SW480 cells

Objective To determine the effect of fibronectin extra type Ⅲ domain A(EDA-FN) on the proliferation and apoptosis of human colorectal cell line SW480.Methods The expression of EDA-FN in SW480 cells was silenced by lentiviral infection.Expression of EDA-FN was examined by Western blot analysis.MTT assay was used to analyze cell viability after infection.Cell apoptosis was analyzed by flow cytometry.The expression of Bax and Bcl-2 were examined by Western blot analysis.Results As compared with the blank control cells(without lentiviral infection) and negative control cells(empty vector infection),the infection of lentiviral inhibited the expression of EDA-FN in SW480 cells.The proliferative capacity of cells was decreased in 24,48,72 and 96 h after lentiviral infection,and all were significantly lower than blank control group and negative control group(P<0.05).The apoptotic rate of SW480 cells after lentiviral infection was(47.76±1.51)%,significantly higher than those of blank control group and negative control group (3.09±1.13)% and(3.71±0.42)% respectively,P<0.05].Western blot analysis showed that the expression of Bax was increased,while that of Bcl-2 was decreased.Conclusion EDA-FN induces the proliferation and inhibits the apoptosis of human colorectal cacer SW480 cells.The inhibitor of EDA-FN may be a new target in the treatment of the malignant tumors.