CAJ | 학술논문

目的探讨唑来膦酸对人鼻咽癌细胞株HNE1增殖、侵袭能力的影响.方法采用四甲基偶氮唑蓝(MTT)法检测细胞毒性,Transwell小室试验检测细胞侵袭能力,酶联免疫吸附试验(ELISA)测定血管内皮细胞生长因子(VEGF)的含量,明胶酶谱法测定基质金属蛋白酶(MMP)2和MMP9的活性,反转录聚合酶链反应(RT-PCR)和Western印迹法测定唑来膦酸对HNE1细胞MMP2、MMP9和VEGF mRNA和蛋白表达水平的影响.结果唑来膦酸浓度为2.5、5、10、20和40μmol/L分别作用HNE1细胞48和72 h,72 h的最高增殖抑制率接近50%,出现在40μmol/L组;但唑来膦酸对HNE1细胞的增殖抑制作用无时间和浓度依赖性.唑来膦酸作用24 h后,0、10、20、40 μmol/L组的穿膜细胞数分别为75.8±2.6、54.8±5.4、44.6±6.4和38.6±8.2,对照组(0 μmol/L组)与处理组比较差异均有统计学意义(均P＜0.01).明胶酶谱法显示40μmol/L组MMP2和MMP9的酶活性明显低,但10、20 μmol/L组无明显变化.ELISA法检测0、10、20、40μmol/L唑来膦酸作用HNE1细胞24h,上清VEGF浓度分别为(5264±89)、(4626±30)、(4155±40)和(1908±171)μg/L,对照组(0 μmol/L组)与处理组比较差异均有统计学意义(均P＜0.01).RT-PCR显示MMP2、MMP9、VEGFmRNA的表达下调,Western印迹法检测其相应蛋白表达水平下降.结论唑来膦酸对人鼻咽癌HNE1细胞的增殖和侵袭能力有抑制作用;唑来膦酸能抑制HNE1细胞VEGF的表达和分泌,抑制MMP2和MMP9酶活性和表达.
목적 탐토서래련산대인비인암세포주HNE1증식、침습능력적영향.방법 채용사갑기우담서람(MTT)법검측세포독성,Transwell소실시험검측세포침습능력,매련면역흡부시험(ELISA)측정혈관내피세포생장인자(VEGF)적함량,명효매보법측정기질금속단백매(MMP)2화MMP9적활성,반전록취합매련반응(RT-PCR)화Western인적법측정서래련산대HNE1세포MMP2、MMP9화VEGF mRNA화단백표체수평적영향.결과 서래련산농도위2.5、5、10、20화40μmol/L분별작용HNE1세포48화72 h,72 h적최고증식억제솔접근50%,출현재40μmol/L조;단서래련산대HNE1세포적증식억제작용무시간화농도의뢰성.서래련산작용24 h후,0、10、20、40 μmol/L조적천막세포수분별위75.8±2.6、54.8±5.4、44.6±6.4화38.6±8.2,대조조(0 μmol/L조)여처리조비교차이균유통계학의의(균P＜0.01).명효매보법현시40μmol/L조MMP2화MMP9적매활성명현저,단10、20 μmol/L조무명현변화.ELISA법검측0、10、20、40μmol/L서래련산작용HNE1세포24h,상청VEGF농도분별위(5264±89)、(4626±30)、(4155±40)화(1908±171)μg/L,대조조(0 μmol/L조)여처리조비교차이균유통계학의의(균P＜0.01).RT-PCR현시MMP2、MMP9、VEGFmRNA적표체하조,Western인적법검측기상응단백표체수평하강.결론 서래련산대인비인암HNE1세포적증식화침습능력유억제작용;서래련산능억제HNE1세포VEGF적표체화분비,억제MMP2화MMP9매활성화표체.
Objective To explore the in vitro anti-tumor effects of zoledronic acid on cell proliferation and invasion in human nasopharyngeal carcinoma cell line HNE1.Methods The cytotoxic effects of zoledronic acid on HNE1 cells were detected by MTT assay, invasion of HNE1 cells by Transwell assay, secretion of (vascular endothelial growth factor)VEGF by (enzyme-linked immunosorbent assay)ELISA and the activities of MMP ( matrix metalloproteinase)2 and MMP9 by gelatine zymography.And the expressions of mRNA and proteins of MMP2, MMP9 and VEGF were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively.Results After a treatment of zoledronic acid at 2.5, 5, 10, 20 and 40 mol/L for 48 h or 72 h, the highest inhibition rate of proliferation at approximately 50% was observed in the 40 mol/L group after 72 h.The inhibitory effect was not in a dose/time-dependent manner.After a 24-hour treatment of zoledronic acid at different concentrations (0, 10, 20 and 40 mol/L), the numbers of membrane-invading cells were 75.8 ±2.6, 54.8 ±5.4, 44.6 ±6.4and 38.6 ± 8.2 respectively (all P ＜ 0.01 ).Gelatinase zymography demonstrated that the activities of MMP2 and MMP9 were inhibited significantly only in cells treated at 0 μmol/L.After a 24-hour exposure to zoledronic acid at O, 10, 20 and 40 μmol/L, the concentrations of VEGF in supernatant were (5264 ± 89 ), (4626 ± 30), (4155 ±40) and ( 1908 ± 171 ) g/L respectively (all P ＜0.01 ).The expressions of mRNA and protein of MMP2, MMP9 and VEGF were down-regulated.Conclusion Zoledronic acid can inhibit the in vitro proliferation and invasion of HNE1 cell through suppressing the secretion of VEGF, the activities of MMP2 and MMP9 and the expressions of VEGF, MMP2 and MMP9.