基质金属蛋白酶2和金属蛋白酶组织抑制因子2在乙酰肝素酶ASODN转染的裸鼠A375细胞移植瘤中的表达

Expressions of matrix metalioproteinase-2 and tissue inhibitor of matrix metalloproteinase-2 in human malignant melanoma transplanted in nude mice

目的 探讨基质金属蛋白酶2（MMP2）和金属蛋白酶组织抑制因子2（TIMP2）在乙酰肝素酶反义寡核苷酸（ASODN）转染的裸鼠恶性黑素瘤移植瘤中的表达.方法 分空白组、无关序列（N-ODN）组、10 μmol/L ASODN组、20 μmol/L ASODN组和30μmol/L ASODN组5组,以脂质体包埋分别转染人恶性黑素瘤细胞株A375,后接种于裸鼠皮下,待成瘤后采用免疫组化SP法检测移植瘤中MMP2和TIMP2蛋白表达的变化.结果 各ASODN组裸鼠移植瘤中MMP2和TIMP2蛋白表达水平均较对照组、N-ODN组显著下调（P＜0.05）；ASODN3个浓度组（10、20、30 μmol/L）相比,差异均有统计学意义（P＜0.05）.结论 乙酰肝素酶ASODN对裸鼠体内恶性黑素瘤移植瘤中的MMP2和TIMP2蛋白的表达有显著抑制效应.

Objective To investigate the expressions of matrix metalloproteinase-2 （MMP2） and tissue inhibitor of MMP2 （TIMP2） in human malignant melanoma transplanted subcutaneously in nude mice.Methods A non-sense oligonucleotide （N-ODN） and an antisense oligodeoxynucleotide （ASODN）against heparanase mRNA were designed and synthesized.Cultured human A375 malignant melanoma cells were classified into 4 groups to be transfected with the N-ODN,ASODN of 10,20,30 μmol/L,respectively,via liposomes.The cells receiving no treatment served as the blank control group.Then,the transfected or untransfected cells were subcutaneously inoculated into nude mice.Six weeks after the transplantation,transplanted tumors were removed from the nude mice and subjected to immunohistochemical staining for the detection of MMP2 and TIMP2 protein expressions.Results The protein expressions of MMP2 and TIMP2 were significantly lower in tumor tissue specimens from nude mice inoculated with ASODN-transfected A375 cells than in those with N-ODN-transfected cells and in those with untransfected cells （P 〈 0.05）,significantly different between the tumor tissue specimens from mice inoculated with A375 cells transfected with 10,20 and 30 μmol/L of ASODN （all P 〈 0.05 ）.Conclusion The ASODN against heparinase displays a marked inhibitory effect on the expressions of MMP2 and TIMP2 proteins in malignant melanoma transplanted in nude mice.