CAJ | 학술논문

目的观察降低中期因子(MK)表达对人前列腺癌PC-3细胞增殖、侵袭及基质金属蛋白酶-9基因(MMP-9)表达的影响.方法用浓度为10 nmol/L的MK基因小干扰RNA(siRNA)转染人前列腺癌PC-3细胞株,应用逆转录-聚合酶链反应(RT-PCR)方法检测MK和MMP-9 mRNA水平,Transwell法和噻唑蓝(MTT)法检测PC-3细胞侵袭和增殖能力,平板克隆形成实验检测PC-3细胞克隆形成能力.结果与对照组比较,MK-siRNA组MK、MMP-9 mRNA水平明显降低(P＜0.05).Transwell实验结果显示,MK-siRNA组穿过滤膜的细胞数为(329.17±10.65)个,明显少于空白对照组[(580.00±10.20)个,P＜0.01]及空载对照组[(575.67 ±11.54)个,P＜0.01].MTT法检测显示,MK-siRNA组细胞增殖受到抑制.平板克隆实验显示:MK-siRNA组克隆形成率为(35.33±2.80)％,低于空白对照组[(71.42±1.11)％,P＜0.01].结论降低MK基因表达能抑制人前列腺癌PC-3细胞的侵袭及增殖,下调MMP-9基因表达,可能是其分子机制之一.
목적 관찰강저중기인자(MK)표체대인전렬선암PC-3세포증식、침습급기질금속단백매-9기인(MMP-9)표체적영향.방법 용농도위10 nmol/L적MK기인소간우RNA(siRNA)전염인전렬선암PC-3세포주,응용역전록-취합매련반응(RT-PCR)방법검측MK화MMP-9 mRNA수평,Transwell법화새서람(MTT)법검측PC-3세포침습화증식능력,평판극륭형성실험검측PC-3세포극륭형성능력.결과 여대조조비교,MK-siRNA조MK、MMP-9 mRNA수평명현강저(P＜0.05).Transwell실험결과현시,MK-siRNA조천과려막적세포수위(329.17±10.65)개,명현소우공백대조조[(580.00±10.20)개,P＜0.01]급공재대조조[(575.67 ±11.54)개,P＜0.01].MTT법검측현시,MK-siRNA조세포증식수도억제.평판극륭실험현시:MK-siRNA조극륭형성솔위(35.33±2.80)％,저우공백대조조[(71.42±1.11)％,P＜0.01].결론 강저MK기인표체능억제인전렬선암PC-3세포적침습급증식,하조MMP-9기인표체,가능시기분자궤제지일.
Objective To observe the effects of down-regulation of midkine (MK) gene expres-sion on proliferation and invasion of prostate cancer PC-3 cells and matrix metalloproteinase (MMP)-9 gene expression.Methods After human prostate cancer PC-3 cells were transfected with midkine small interfer-ing RNA (siRNA) at the concentration of 10 nmol/L,the mRNA expression levels of midkine and MMP-9 were examined by reverse transcription-polymerase chain reaction (RT-PCR).The invasion and prolifera-tion ability was determined by Transwell and methyl thiazol tetrazolium (MTT) assay respectively.The PC-3 cell clony formation ability was evaluated by plate colony formation assay.Results As compared with the control group,the mRNA expression levels of midkine and MMP-9 gene were greatly inhibited in pros-tate cancer PC-3 cells transfected with MK-siRNA (P ＜ 0.05).Transwell assay showed that the number of PC-3 cells penetrating the membrane in the siRNA group (329.17 ± 10.65) was greatly decreased as com-pared with the Con-A group (580.00 ± 10.20,P ＜ 0.01) and Con-B group (575.67 ± 11.54,P ＜ 0.01).MTT assay showed that the cell proliferation was significantly inhibited in MK-siRNA group.The result of plate colony formation assays showed that the colony formation rate in the siRNA group [(35.33 ± 2.80) ％] was significantly decreased as compared with Con-A group [(71.42 ± 1.11) ％] (P ＜ 0.01).Conclusion Down-regulation of midkine gene can inhibit the invasion and proliferation of prostate cancer cells through down-regulation of MMP-9 gene.