摘要
目的观察乙酰肝素酶反义寡核苷酸(ASODN)对人恶性黑素瘤A375细胞增殖及乙酰肝素酶蛋白表达的影响。方法以空白组和无关序列(N-ODN)组为对照,设计、合成4条ASODN链(ASODN-t1,t2,t3和t4),脂质体包埋后分别以10μmol/L、20μmol/L、30μmol/LN-ODN和ASODN转染A375细胞,台盼蓝排斥试验检测细胞增殖情况,免疫组织化学法检测乙酰肝素酶蛋白表达的变化。结果转染72h后,各ASODN组A375细胞计数较对照组、N-ODN组均减少(P均=0.000);每条链的10μmol/L、20μmol/L、30μmol/L3个浓度组相比,差异均有统计学意义(P均=0.000);30μmol/LASODN-t2组与其他各组相比,差异均有统计学意义(P均<0.05)。转染48h后,30μmol/LASODN-t2组乙酰肝素酶蛋白的表达较对照组、N-ODN组下调(P均=0.000)。结论乙酰肝素酶ASODN可抑制A375细胞增殖,下调乙酰肝素酶蛋白的表达。
Aim：To study the effects of heparinase antisense oligodeoxynucleotide （ASODN） on the proliferation of A375 cells and the expression of heparinase protein in A375 cells. Methods： tleparinase ASODN of fuur different sequences （ASODN-tl, t2, t3, and t4 ） and its controlled nonsense oligodeoxynucleutide （ N-ODN ） were designed and synthesized. The ODNs with the final concentration of 10 μmol/L, 20μmol/L, and 30 μmol/L were delivered into A375 cells by Oligofectamine^TM reagent. The viability of the cells was determined by the trypanblue exclusion test. Heparinase protein was detected by immunohistochemistry. Results ： At 72 h after incubation, the cell number exposed to hepariuase ASODN were all decreased compared with the control group and N-ODN group（ P = 0. 000）. For every sequence of ASODN, there was all significant difference between the cell number of 10 μmol/L and 20 μmol/L ASODN, 20 μmol/L and 30 μmol/L ASODN, or 10 μmol/L and 30 μmol/L ASODN（P =0.000）. The cell number in 30 μmol/L ASODN-t2 group was lower than those of any other groups （ P 〈 0.05 ）. At 48 h after incubation, the expression of heparinase protein was significantly lower in the ASODN groups than that in N-ODN group as well as that in the cnntrol group（ P = 0. 000）. Conclusion ： Hepa- rinase ASODN could inhibit the cell proliferation and down-regulate the expression of heparinase protein in A375 cells.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2006年第6期1067-1070,共4页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省医学科技攻关计划基金资助项目20060017
