miRNA-221 promotes cutaneous squamous cell carcinoma progression by targeting PTEN.

BACKGROUND: Cutaneous squamous cell carcinoma (CSCC) is a common type of skin malignancy. MicroRNA-221 (miRNA-221) is a critical non-coding RNA in tumor initiation and progression. However, the molecular mechanisms of miRNA-221 in the development of CSCC remain unknown. This study investigated the expression of miRNA-221 in CSCC and its potential tumor biological functions. METHODS: MTT assay, colony assay, PCR, and Western blot were adopted. RESULTS: In this study, miRNA-221 expression was significantly higher in CSCC tissues and cell lines than in normal tissues and cells (P < 0.05). Further functional experiments indicated that miRNA-221 knockdown inhibited the proliferation and cell cycle, while upregulation of miRNA-221 presented the opposite role. The dual reporter gene assays indicated that PTEN is a direct target gene of miRNA-221. PTEN protein or mRNA levels were decreased after the cells were transfected with miR-221 mimics. CONCLUSIONS: Taken together, the obtained results indicated that miR-221 plays an oncogenic function in CSCC by targeting PTEN and further suggest that miR-221 may be a potential target for CSCC diagnosis and treatment.

Bromodomain protein BRD4 promotes cell proliferation in skin squamous cell carcinoma.

The present study examined the expression and biological functions of bromodomain-containing protein 4 (BRD4) in skin squamous cell carcinoma (SCC) cells. Our results show that BRD4 mRNA and protein expression was upregulated in human skin SCC cells, as compared to its level in the normal skin keratinocytes and fibroblasts. Treatment with BRD4 inhibitors, JQ1 and CPI203, resulted in proliferation inhibition, apoptosis and cell cycle arrest in both established (A431 cell line) and primary skin SCC cells. Furthermore, BRD4 knockdown (by targeted shRNAs) or knockout (by CRISPR/Cas9) largely inhibited A431 cell proliferation. Reversely, forced-overexpression of BRD4 in A431 cells facilitated cell proliferation. We show that BRD4 is required for the expression of several oncogenes, including cyclin D1, Bcl-2 and MYC. BRD4 inhibition, knockdown or knockout significantly decreased above oncogene expression in SCC cells. In vivo, CRISPR/Cas9-mediated BRD4 knockout significantly suppressed A431 xenograft tumor growth in severe combined immunodeficient (SCID) mice. Together, our results suggest that BRD4 could be a novel and pivotal oncogenic protein of skin SCC.

Association of plasminogen activator inhibitor-1 and vitamin D receptor expression with the risk of keloid disease in a Chinese population.

Keloid disease (KD) is a benign fibroproliferative scarring condition of unknown etiopathogenesis. Plasminogen activator inhibitor-1 (PAI-1) and vitamin D receptor (VDR) have been shown to play important roles in the progression of tissue fibrosis; therefore, both these genes are potential susceptibility genes for KD. We aimed to determine whether the gene expression levels of PAI-1 and VDR are altered in Chinese KD patients. We measured the expression of PAI and VDR in human peripheral blood lymphocytes in 236 patients with keloid and 219 age- and sex-matched healthy controls by quantitative real-time polymerase chain reaction. We found that PAI-1 expression in peripheral blood lymphocytes was significantly higher in patients with KD than in control individuals (p < 0.0001), while VDR expression was significantly lower in KD patients than in control individuals (p < 0.0001). High levels of PAI-1 and low levels of VDR expression were significantly associated with an increased risk for KD. PAI-1 and VDR might play important roles in keloid development. Gene expression levels of PAI-1 and VDR may, therefore, be used as potential markers for the prediction of keloid development after scarring.

Impact of human leukocyte antigen matching and recipients' panel reactive antibodies on two-year outcome in presensitized renal allograft recipients.

BACKGROUND: Renal transplantation in sensitized candidates remains a highly significant challenge worldwide. The production of panel reactive antibody (PRA) against human leukocyte antigen (HLA) is a major risk factor in presensitized recipients. The aim of this study was to evaluate the impact of HLA matching and recipients' PRA on two-year outcome in presensitized renal allograft recipients. METHODS: We determined the percentage of panel reactivity and specificity of anti-HLA immunoglobulin (Ig) G antibodies in 73 presensitized renal allograft recipients compared with 81 unsensitized recipients (control group). HLA genotyping of both recipients and corresponding donors was performed by PCR with sequence-specific primers (PCR-SSP). We analyzed the factors influencing the early graft outcome (two-year rejection rates and survival rates of the grafts), including HLA mismatching, class and degree of panel reactivity, and target antigen of donors. RESULTS: Presensitized recipients had a worse two-year outcome than unsensitized recipients (P = 0.019 for rejection rate, P = 0.01 for survival rate). The difference in number of HLA-mismatched alleles with either 6-antigen matching (Ag M) standard or amino acid residue matching (Res M) standard was not significant between the rejection and non-rejection groups of presensitized recipients or between the graft survival group and graft loss group. Compared with the control group, recipients with both PRA-I and PRA-II antibodies had a significantly worse two-year outcome (P = 0.001 for rejection rate, P = 0.002 for survival rate). The two-year outcomes of the peak PRA >/= 50% group and its subgroup, at-transplant PRA > or = 50% group, were significantly worse compared with the control group (P = 0.025 and P = 0.001 for rejection rate, P = 0.043 and P = 0.024 for survival rate). The rejection rates of the at-transplant target antigen positive group and its subgroup, HLA-I target antigen positive group, were significantly higher than the control group (P = 0.001 and P = 0.001), target antigen negative group (P = 0.003 and P = 0.001), and peak target antigen positive with negative at-transplant target antigen group (P = 0.024 and P = 0.002). Two-year graft survival rates of the target antigen positive group and HLA-I target antigen positive group were significantly lower than the control group (P = 0.012 and P = 0.001). The two-year outcome of target antigen unknown group was similar to that of the target antigen positive group. Presensitized recipients with pre-transplant plasmapheresis or immunoadsorption (PRA prepared group) had a better but non-significant two-year outcome than the control group. However, the PRA unprepared presensitized recipients were different to the control group (P = 0.004 for rejection rate and P = 0.005 for survival rate). Hyperacute rejection (HR) occurred in three recipients with positive HLA-I target antigen and without mismatch according to Res M and in one case with positive PRA-II (for an unknown target antigen). No HR occurred in eight cases with positive HLA-II target antigens. CONCLUSIONS: Pre-transplant PRA preparations might improve the access of presensitized patients to renal donors. Avoiding antigen-positive donors remains a fundamental measure in preventing HR and early rejections.

Impact of human leukocyte antigen matching and recipients' panel reactive antibodies on two-year outcome in presensitized renal allograft recipients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Renal transplantation in sensitized candidates remains a highly significant challenge worldwide. The production of panel reactive antibody (PRA) against human leukocyte antigen (HLA) is a major risk factor in presensitized recipients. The aim of this study was to evaluate the impact of HLA matching and recipients' PRA on two-year outcome in presensitized renal allograft recipients.</p><p><b>METHODS</b>We determined the percentage of panel reactivity and specificity of anti-HLA immunoglobulin (Ig) G antibodies in 73 presensitized renal allograft recipients compared with 81 unsensitized recipients (control group). HLA genotyping of both recipients and corresponding donors was performed by PCR with sequence-specific primers (PCR-SSP). We analyzed the factors influencing the early graft outcome (two-year rejection rates and survival rates of the grafts), including HLA mismatching, class and degree of panel reactivity, and target antigen of donors.</p><p><b>RESULTS</b>Presensitized recipients had a worse two-year outcome than unsensitized recipients (P = 0.019 for rejection rate, P = 0.01 for survival rate). The difference in number of HLA-mismatched alleles with either 6-antigen matching (Ag M) standard or amino acid residue matching (Res M) standard was not significant between the rejection and non-rejection groups of presensitized recipients or between the graft survival group and graft loss group. Compared with the control group, recipients with both PRA-I and PRA-II antibodies had a significantly worse two-year outcome (P = 0.001 for rejection rate, P = 0.002 for survival rate). The two-year outcomes of the peak PRA >/= 50% group and its subgroup, at-transplant PRA > or = 50% group, were significantly worse compared with the control group (P = 0.025 and P = 0.001 for rejection rate, P = 0.043 and P = 0.024 for survival rate). The rejection rates of the at-transplant target antigen positive group and its subgroup, HLA-I target antigen positive group, were significantly higher than the control group (P = 0.001 and P = 0.001), target antigen negative group (P = 0.003 and P = 0.001), and peak target antigen positive with negative at-transplant target antigen group (P = 0.024 and P = 0.002). Two-year graft survival rates of the target antigen positive group and HLA-I target antigen positive group were significantly lower than the control group (P = 0.012 and P = 0.001). The two-year outcome of target antigen unknown group was similar to that of the target antigen positive group. Presensitized recipients with pre-transplant plasmapheresis or immunoadsorption (PRA prepared group) had a better but non-significant two-year outcome than the control group. However, the PRA unprepared presensitized recipients were different to the control group (P = 0.004 for rejection rate and P = 0.005 for survival rate). Hyperacute rejection (HR) occurred in three recipients with positive HLA-I target antigen and without mismatch according to Res M and in one case with positive PRA-II (for an unknown target antigen). No HR occurred in eight cases with positive HLA-II target antigens.</p><p><b>CONCLUSIONS</b>Pre-transplant PRA preparations might improve the access of presensitized patients to renal donors. Avoiding antigen-positive donors remains a fundamental measure in preventing HR and early rejections.</p>