ABSTRACT
ObjectiveTo investigate the expressions ofheparinase,matrix metalloproteinase 2 (MMP2) and tissue inhibitor of metalloproteinase 2(TIMP2) in malignant melanoma lesions and their significance.MethodsSkin specimens were obtained from the lesions of 30 patients with malignant melanoma,30 patients with melanocytic nevus and the normal skin of 15 healthy controls.Immunohistochemical SP method was used to detect the protein expression of heparinase,MMP2 and TIMP2.ResultsThe malignant melanoma tissue specimens significantly differed from the melanocytic nevus and control tissue specimens in the expression rate of heparinase (63.33％ vs.6.67％ and 0.00,x2 =21.172,27.805,both P ＜ 0.01 ),MMP2 (70.00％ vs.13.33％ and 0.00,x2 =19.817,19.866,both P＜ 0.01) and TIMP2(60.00％ vs.6.67％ and 0.00,x2 =19.200,15.000,both P ＜ 0.01 ).ConclusionThe expression of heparinase,MMP2 and TIMP2 is significantly higher in malignant melanoma lesions than in melanocytic nevus lesions and normal skin tissue.
ABSTRACT
ObjectiveTo 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.ResultsThe 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 ).ConclusionThe ASODN against heparinase displays a marked inhibitory effect on the expressions of MMP2 and TIMP2 proteins in malignant melanoma transplanted in nude mice.
ABSTRACT
Objective To study the relationship of matrix metalloproteinase 7(MMP7)expression to the infiltration and metastasis of malignant melanoma.Methods Various concentrations(10,20,30 μmol/L)of heparanase antisense oligodeoxynucleotide (ASODN)was used to transfect human malignant melanoma cell line A375.Then,the transfected cells were subcutaneously inoculated into the right back of nude mice.Six weeks after the inoculation,transplanted tumors were isolated from the mice,and the levels of MMP7 protein expression were assessed by SP immunohistochemistry.The expression rate of MMP7 was also examined by SP immunohistochemistry in A375 cells and in tissue samples from patients with malignant melanoma(n=30),iunctional nevus(n=30)and normal human controls(n=15).Results The expression rate of MMP-7 was 83.33%(25/30),6.67%(2/30)and 0(0/15)respectively in malignant melanoma,iunctional nevus and normal skin tissue samples;there was a significant difierence in the positivity rate of MMP-7 between the malignant melanoma group and iunctional nevus group (x2=35.62,P=0.000)as well as normal control group(x2=28.12,P=0.000),but not between the junctional nevus group and normal control group(P＞0.05).The expression of MMP-7 in transplanted tumor was decreased to a varying degreee by three levels of ASODN,and the inhibitory effects of 30 mol/L ASODN were the strongest(P＜0.05).MMP-7 protein was also highly expressed in A375 cells.Conclusions The expression of MMP-7 in cutaneous malignant melanoma is higher than that in iunctional nevus and normal skin tissue.Hpa-ASPDN significantly inhibits the expression of MMP-7 in transplanted tumors in nude mice.MMP-7 is highly expressed in A375 cells.
ABSTRACT
To investigate the gene mutation in a pedigree with X-linked ichthyosis (XLI) and to explore the relationship between the mutation and its clinical manifestations, genomic DNA of affected members, the normal member of the pedigree and 50 unrelated normal members was extracted with a whole blood genomic DNA extraction kit and the DNA was used as a template for the polymerase chain reaction (PCR)-mediated amplification of exon 1 and exon 10 of the STS gene. hHb6 (human hair basic keratin) gene was used as the internal control. Our results showed that the STS gene was deleted in affected members in the pedigree with X-linked ichthyosis. The normal member of the pedigree and 50 unrelated normal members had no such deletion. The proband and his mother had products in the internal control after PCR amplification. The blank control had no product. It is concluded that deletion of the STS gene existed in this pedigree with X-linked ichthyosis, and it is responsible for the unique skin lesions of X-linked ichthyosis.
ABSTRACT
Objective To investigate the effects of tazarotene on the proliferation of cultured human keratinocytes and IFN-?-induced expression of HLA-DR in those cells.Methods Keratinocytes were cul-tured from normal human skin in vitro,and were treated with various concentrations of tazarotene(10 -5 ,10 -6 ,10 -7 mol/L).At24h,48h after treatment,the effects on cell proliferation were assessed by MTT method.The expression of HLA-DR was determined using immunocytochemistry techniques in cultured human ker-atinocytes incubated with tazarotene,IFN-?or both for24h.Results①The proliferation of keratinocytes was decreased when exposed to10 -7 -10 -5 of tazarotene as compared to non-exposed keratinocytes after24h and48h.Moreover,the effects on cell proliferation by tazarotene were dose-dependent;②There was rare expression of HLA-DR in normal human keratinocytes.③HLA-DR expression was inducible significantly with500u/mL of IFN-?,but failed to be induced with10 -6 mol/L of tazarotene,in keratinocytes at24h af-ter treatment.④After24h combined treatment of10 -7 -10 -5 mol/L of tazarotene and IFN-?,the induction of HLA-DR expression was significantly stronger,in a dose-dependent manner,than IFN-?alone(P