微RNAs-107对胰腺癌PANC-1细胞增殖、衰老和侵袭的影响

The effects of miRNAs-107 on cell proliferation, senescence and invasion of pancreatic cancer PANC-1

目的探讨特异性微RNAs（miR）-107对胰腺癌PANC-1细胞增殖、衰老及侵袭的影响。方法应用实时荧光定量PCR（qRT—PCR）检测3种人胰腺癌细胞系PANC-1、ASPC-1、BXPC-3和正常胰腺HTERT—HPNE细胞中miR-107表达情况。通过qRT—PCR观察50nmol／Lanti-miR-107对胰腺癌PANC-1细胞miR-107表达的影响；通过M，ITr法观察anti—miR-107对胰腺癌PANC-1细胞增殖的影响；通过B-半乳糖苷酶染色检测anti-miR-107对胰腺癌细胞衰老的影响；通过transwell法测定anti—miR-107对胰腺癌细胞侵袭的影响，应用qRT—PCR检测anti—miR-107对PCNA、P16INK4A和MMP2表达的影响。结果miR-107在3种胰腺癌细胞中的表达明显高于正常对照细胞（P〈0．01）。转染anti—miR-107后，PANC-1细胞增殖受抑，衰老率明显增加（P〈0．05），但侵袭率未受明显影响。转染anti—miR-107后PCNA下调，P16INK4A表达明显增加，MMP2变化不明显。结论miR-107可能部分通过影响PCNA和P16INK4A的表达促进胰腺癌细胞的增殖和逃避细胞衰老，对侵袭影响不明显。miR-107有可能成为胰腺癌生物治疗的新靶点。

Objective To investigate the effects of miRNAs-107 （ miR-107 ） on pancreatic cancer proliferation, senescence and invasion. Methods MiR-107 expression levels in 3 pancreatic cancer cell lines PANC-1, ASPC-1, BXPC-3 and normal pancreatic HTERT-HPNE cells were studied by quantitative reverse transeription-polymerase chain reaction （qRT-PCR）. PANC-1 cells were transfected with 50 nmol/L anti-miR-107 or negative control using Lipofectamine 2000. After transfection, the miR-107 expression was measured by qRT-PCR. Cell proliferation was tested by methylthiazol tetrazolium （MTT） assay. Cell senes- cence was detected by B-galactosidase staining. The expression levels of PCNA, P16INK4A and MMP2 were measured by qRT-PCR. Results Compared with the HTERT-HPNE cells, the expression level of miR-107 in 3 pancreatic cancer cell lines was significantly increased （ P 〈 0.01 ）. After transfected with 50 nmol/L anti-miR-107, cell proliferation was inhibited, and cell senescence were increased in PANC-1 cells （ P 〈 0. 05）, and there was no obvious change in cell invasion. Compared with the HTERT-HPNE cells, after transfected with anti-miR-107, the PCNA expression was significantly decreased and P16INK4A was signifi- cantly increased, but expression of MMP2 didn＇ t change significantly. Conclusions These results demon- strate that miR-107 promotes the proliferation and escapes cell senescence in PANC-1 cells by targeting PCNA and P16INK4A. But it has no obvious effects on cell invasion. Therefore, it may be a new target for the biologic therapy for pancreatic cancer.