FHL1对肺腺癌细胞株A549生物学特性的影响

 目的 研究FHL1的表达对人肺腺癌细胞株A549生物学特征的影响。方法 以FHL1 mRNA及shRNA慢病毒载体转染肺腺癌细胞株A549,并采用Western blot及RT-PCR法筛选检测稳定过表达FHL1肺腺癌细胞株模型及低表达细胞株模型;运用CCK-8实验、Transwell实验、流式细胞仪、细胞集落形成实验等方法对其增殖、侵袭、凋亡及锚定非依赖性生长生物学特性进行检测评估。结果 FHL1稳定过表达及低表达组的Western blot及RT-PCR法检测显示病毒转染效果可靠稳定;CCK-8实验提示低表达组的细胞增殖情况明显高于过表达组及空白组(P=0.002 2);流式细胞仪检测提示过表达组的细胞总体凋亡率为50.48%,明显高于两个低表达组(16.41%、20.12%)及空白对照组(25.90%),差异有统计学意义(P<0.001);Transwell实验提示过表达组细胞表现出的侵袭性明显低于低表达组及空白组差异有统计学意义(P<0.001);成集落实验显示过表达组细胞株形成的集落数为28.7±6.0,明显少于两个低表达组(157.0±6.4、150.7±9.5),差异有统计学意义(P<0.001)。结论 稳定过表达FHL1肺腺癌细胞株A549模型及低表达模型构建成功。FHL1的表达对于肺腺癌细胞株A549的增殖、侵袭及转移均有抑制作用,并能够诱导其加速凋亡。

Effects of FHL1 on biological character of lung adenocarcinoma cell line A549

Objective To explore the effect of expression of FHL1 on biological character of lung adenocarcinoma cell line A549. Methods Lung adenocarcinoma cell line A549 was transfected by FHL1 mRNA and shRNA lentivirus vector. The stable overexpression of FHL1 in lung adenocarcinoma cell line A549 model and low expression of cell line model were tested by Western blot and RT-PCR method.CCK-8 test, Transwell invasion, flow cytometry and Colony-forming Assay were used to test the proliferation, invasion, apoptosis and anchorage independence growth of the cell line. Results The difference between the stable overexpression of FHL1 group and its low expression group was obvious tested by Western blot and RT-PCR. CCK-8 test showed higher cell proliferation of the low expression group than that of the overexpression group and the blank group (P=0.002 2). Flow cytometry showed that the apoptosis rate of the overexpression group was 50.48%, significantly higher than that of the two low expression groups (16.41%, 20.12%) and blank group (25.90%, P<0.001). The invasiveness of the overexpression group was obviously lower than the low expression group and blank group in the Transwell invasion (P<0.001). Colony-forming Assay showed that the colony number of the overexpression group was 28.7±6.0, significantly less than the two low expression groups (157.0±6.4, 150.7±9.5, P<0.000 1). Conclusions The stable overexpression and low expression of FHL1 in lung cancer cell line A549 models were successfully constructed. The overexpression of FHL1 inhibited the proliferation, invasion and metastasis, and accelerated apoptosis in lung adenocarcinoma cell line A549.