李玉花 刘希光 张红军 宋浩 徐明金 姜韬 肖文静 赵淑芬 于晓芸.成纤维细胞生长因子3-siRNA对肺癌细胞A549侵袭性和基质金属蛋白
酶9 表达的影响[J].,2015,15(22):4235-4238 |
成纤维细胞生长因子3-siRNA对肺癌细胞A549侵袭性和基质金属蛋白
酶9 表达的影响 |
FGFR3- siRNA Impacts on Lung Cancer A549 Cells Invasivenessand MMP9 Expression |
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DOI: |
中文关键词: 肺癌 siRNA MMP9 FGFR3 |
英文关键词: Lung cancer siRNA MMP9 FGFR3 |
基金项目:山东省自然科学基金项目(320.6750.13210);卫生部科研课题(W2012FZ078) |
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中文摘要: |
目的:探讨人成纤维细胞生长因子3(Fibroblast growth factor3,FGFR3)基因沉默对人类肺腺癌A549 细胞侵袭能力及其对基
质金属蛋白酶9(Matrix metaloproteinases 9,MMP9)基因表达的影响。方法:细胞分为3 组:A 组:实验组,即FGFR3 特异性小干扰
RNA(Small interfering RNA, siRNA)(siRNA-FGFR3)干扰组;B 组:阴性对照组,即FGFR3 非特异性阴性对照siRNA(siRNA-NC)
干扰组;C 组:空白对照组,无siRNA 干扰;通过核酸转染试剂脂质体LipofectamineTM2000(Lipo2000)转染A549细胞;倒置荧光
显微镜观察Lipo2000 转染效率;转染后A549 细胞的侵袭能力用Transwell 实验检测;实时荧光定量聚合酶链反应(Real-time
quantitative polymerase chain reaction, Real-time PCR)用于检测转染前后FGFR3 及MMP9 mRNA的表达水平。结果:Lipo2000 介
导的FAM-siRNA对肺腺癌A549 细胞的转染效率可达80%;在转染36 h后,Transwell 实验结果显示A组较B组、C 组侵袭能力
显著降低(P<0.01)。Real-time PCR 结果显示,A组较B、C 组的FGFR3 和MMP9 基因表达量明显下调(P<0.01)。结论:FGFR3 基
因沉默可明显抑制肺腺癌A549 细胞的侵袭能力,并能下调MMP9表达。为肺癌的治疗提供了新的靶点。 |
英文摘要: |
Objective:In order to explore the human fibroblast growth factor 3 (FGFR3) gene silencing impact on the invasive
ability of human lung adenocarcinoma A549 cells and matrix metalloproteinase-9 (MMP9) gene expression.Methods:A549 cells were
classified into three groups: A: the experimental group, siRNA inhibited FGFR3 specifically siRNA-FGFR3 interference group; group B:
negative control group, siRNA-NC interference group C: blank control group, no siRNA interference group. A549 cells were transfected
with nucleic acid transfection reagent liposomes LipofectamineTM2000 (Lipo2000). The transfection efficiency was observed under the
inverted fluorescence microscope. The invasive ability of A549 cells was detected by the transwell assay after transfection. The
expression of FGFR3 and MMP9 mRNA before and after transfection were measured by Real-time polymerase chain reaction (Real-time
PCR).Results:The transfection efficiency with Lipo2000 mediated FAM-siRNA to A549 cells was up to 80% under the inverted
fluorescence microscope. Transwell experiments showed the invasive capability inhibited in A group than in B and C group after 36h
transfection significantly (P <0.01). Real-time PCR showed that the expression of FGFR3 and MMP9 were decreased in A group
compared with B and C group significantly (P<0.01).Conclusion:Experimental findings indicated that FGFR3 gene silencing can
inhibit the invasion ability of A549 cells and down-regulate the expression of MMP9 significantly. A new target for the treatment of lung
carcinoma is supplied by this work. |
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