LOX基因降表达对人阴道壁成纤维细胞中ELN及MMP-2基因表达的影响

Influence of LOX gene drop expression in human vaginal wall fibroblasts on the mRNA expressions of ELN and MMP-2

目的研究赖氨酰氧化酶(LOX)基因对人阴道壁成纤维细胞中弹性蛋白(ELN)、基质金属蛋白酶-2(MMP-2)基因mRNA表达的影响。方法将原代培养的人阴道壁成纤维细胞分成3个组(LOX基因降表达组、阴性对照组、空白对照组),分别用LOX-sh RNA重组慢病毒、阴性对照慢病毒转染前2个组细胞,计算转染效率。流式细胞仪分选慢病毒感染后带绿色荧光的人阴道壁成纤维细胞,传代后收集部分细胞,通过实时荧光定量聚合酶链反应(PCR)检测3个组成纤维细胞中LOX、ELN、MMP-2的mRNA表达水平。结果 LOX-sh RNA慢病毒转染人阴道壁成纤维细胞共8例(绝经前4例,绝经后4例)。LOX基因降表达组成纤维细胞胞体变小,生长变慢,贴壁性降低,LOX mRNA的表达水平较阴性对照组和空白对照组显著下降(P<0.05)。LOX基因降表达组ELN mRNA表达较对照组明显降低,而MMP-2 mRNA表达较对照组明显升高。阴性对照组和空白对照组的LOX mRNA、ELN mRNA、MMP-2 mRNA表达差异无统计学意义(P>0.05)。结论 LOX基因降表达后人阴道壁成纤维细胞ELN mRNA表达显著降低,MMP-2 mRNA表达显著增高,提示LOX通过调节ELN和MMP-2的表达参与盆腔脏器脱垂(POP)的发生。

Objective To study the influence of lysyl oxidase （ LOX） gene drop expression in human vaginal wall fibroblasts on the mRNA expressions of elastin （ELN） and matrix metalloproteinase-2 （MMP-2 ） . M e th o d s The primary-cultured human vaginal wall fibroblasts were classified into 3 groups,LOX gene drop expression group, negative control group and blank control group. LOX-shRNA lentiviruses were transfected into the fibroblasts of LO^ X gene drop expression group. Negative control lentiviruses were transfected into the fibroblasts of negative control group. The transfection efficiency was calculated. Flow cytometry sorted fibroblasts with green fluorescence,and some cells were collected and determined. Real-time fluorescence quantitation polymerase chain reaction （PCR） was used to determine the mRNA expressions of LOX,ELN and MMP-2. R e su l t s A total of 8 samples were expressed in the fibroblasts in LOX gene drop expression group, including 4 cases of premenopausal women and 4 cases of postmenopausal women. The fibroblasts in LOX gene drop expression group became smaller, the growth was slow, and the adherence decreased. The expression of LOX mRNA in LOX gene drop expression group was lower than those in negative and blank control groups （P〈0.05） . The expression of ELN mRNA in LOX gene drop expression group was lower than those in negative and blank control groups,while the expression of MMP-2 mRNA was higher. There was no statistical significance for LOX mRNA, mRNAand MMP-2 mRNA expressions between negative and blank control groups （P〉0.05 ） . Conclusions The expression of ELN mRNA decreases, however, that ofMWP-2 mRNA increased. LOK may participate in the occurrence of pelvic organ prolapse （POP） by regulating the expressions of ELN mRNA and MMP-2 mRNA.