RESUMEN
Keloid is a disorder of fibroproliferative diseases that occurs in wounds, characterized by an exaggerated response to injury. The key factor responsible for the disease process has not been identified. This study sought to elucidate the role of ß-catenin in the regulation of keloid phenotypes and signaling. Expression of ß-catenin in keloid and normal non-keloid samples was measured by real-time polymerase chain reaction. Knockdown of ß-catenin was achieved by delivering small interfering RNA to target ß-catenin. Cell proliferation, cell cycle progression, and apoptosis of keloid cells were measured by functional assays in vitro. The proteins related to keloid fibrosis were measured by Western blotting. ß-catenin expression was significantly upregulated in keloid tissue samples compared with the normal non-keloid age-adjusted skin sample counterparts. Functionally, targeting ß-catenin with lipofection-delivered small interfering RNA oligonucleotide inhibited the proliferation and cell cycle arrest in G0/G1 phase and increased apoptosis of fibroblast cells, accompanied by downregulation of Wnt2 and cyclin D1 as well as the phosphorylation level of glycogen synthase kinase 3 beta in the keloid fibrosis. Our study supports a crucial role of ß-catenin in the regulation of fibroproliferation and extracellular matrix deposition. Targeting ß-catenin using small interfering RNA oligonucleotide may be a promising approach for preventing excessive fibroproliferative development after wound healing and may lead to the development of novel strategies for restoring keloid diseases.
Asunto(s)
Proliferación Celular/genética , Queloide/genética , Proteína wnt2/genética , beta Catenina/biosíntesis , Apoptosis/genética , Ciclina D1/genética , Femenino , Fibroblastos/metabolismo , Fibroblastos/patología , Regulación Neoplásica de la Expresión Génica , Glucógeno Sintasa Quinasa 3 beta/genética , Humanos , Queloide/patología , Fosforilación , ARN Interferente Pequeño/genética , Transducción de Señal/genética , beta Catenina/genéticaRESUMEN
To study the effect of knocking down wingless-related MMTV integration site 2 (Wnt2) expression by RNAi on the growth and signaling pathways of ex vitro-cultured keloid fibroblasts (KFB).Human KFB were isolated from 10 keloid patient specimens. The KFB cells were then transfected with 4 pairs of small interfering RNA (siRNA) targeting human Wnt2, respectively. Reverse transcriptase-polymerase chain reaction and Western blot analysis were conducted to verify the knock down of Wnt2, and the expression of ß-catenin glycogen synthase kinase-3ß (GSK-3ß) and cyclin D1 were examined.siRNA Wnt2 transfection (siWnt2) resulted in the significant inhibition of Wnt2 expression at both the mRNA and protein levels. The expression of ß-catenin, GSK-3ß, p-GSK-3ß, and cyclin D1 at the protein level also decreased in siWnt2 cells. siWnt2 resulted in a substantially slower growth and significant delay in cell doubling time of the KFB cells compared with control groups. Further, the siRNA knock down of GSK-3ß and ß-catenin resulted in slower proliferation rates, respectively.Wnt2 siRNA has an inhibitive effect on keloid fibroblast proliferation, which may be a potential therapeutic approach for keloid and other human fibrotic diseases.
Asunto(s)
Fibroblastos/metabolismo , Queloide/metabolismo , Interferencia de ARN/fisiología , Transducción de Señal/fisiología , Proteína wnt2/biosíntesis , Adolescente , Adulto , Western Blotting , Niño , Preescolar , Ciclina D1/biosíntesis , Femenino , Glucógeno Sintasa Quinasas/biosíntesis , Humanos , Masculino , Persona de Mediana Edad , ARN Interferente Pequeño , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Adulto JovenRESUMEN
AIM: To study the SSTR1, 2, 3, 4, 5 expression and their relationships with clinico-pathological factors, cell proliferation, Bcl-2 and p53 expression in colorectal cancer cells. METHODS: Immunohistochemical staining of five SSTR subtypes, Ki-67, Bcl-2 and p53 was performed by the standard streptavidin-peroxidase (SP) technique for the paraffin sections of 127 colorectal cancers. and expression of five SSTR subtypes in 40 specimens of normal colorectal mucosae was detected with the same method. RESULTS: Positive staining for five SSTR subtypes was observed in colorectal cancer cells and normal colorectal mucosae. SSTR1 was the most predominant subtype in both colorectal cancer and normal colorectal mucosa, and the second was SSTR5 or SSTR2. As compared with normal colorectal mucosa, SSTR4 was more frequently expressed in colorectal cancer cells (2.5% vs 18.9%, P<0.05); the expression of SSTR2, 4, 5 in moderately to well differentiated colorectal adenocarcinoma was significantly higher than that in poorly differentiated ones (P<0.05), the SSTR1 expression in colorectal cancer with positive lymph node metastasis was significantly higher than that with negative lymph node metastasis (72.2% and 54.5%,P<0.05). In addition, in the ulcerative type of colorectal cancer, SSTR2 expression was obviously decreased (P<0.05); the correlation did not reach a statistical significance between the five SSTR subtypes expression and Dukes'stages (P>0.05), but the frequency of SSTR1 expression increased with Dukes'stage, while SSTR3 and SSTR5 expression decreased with Dukes' stage. Moreover, there was no correlation between expression of the five SSTR subtypes and other clinicopathological factors such as age, sex, tumor site, tumor depth, distant metastasis. The proliferative indexes in colorectal cancer cells with negative expression of SSTR2 and SSTR3 were significantly higher than that with positive expression (P<0.05). The Bcl-2 expression in colorectal cancer cells with positive expression of SSTR1, 2, 3, 5 was significantly lower than that with negative expression (P<0.05). There was no correlation between five SSTR subtypes and p53 expression. CONCLUSION: The most predominant SSTR subtype is SSTR1, and the second is SSTR2 or SSTR5. Five SSTR subtypes play different roles in the development of colorectal cancer. SSTR2 and SSTR3 can inhibit the proliferation and promote apoptosis of tumor cells.
Asunto(s)
Neoplasias Colorrectales/química , Neoplasias Colorrectales/patología , Antígeno Ki-67/análisis , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Receptores de Somatostatina/análisis , Proteína p53 Supresora de Tumor/análisis , Adulto , Anciano , Anciano de 80 o más Años , Proliferación Celular , Femenino , Humanos , Inmunohistoquímica , Masculino , Proteínas de la Membrana/análisis , Persona de Mediana Edad , Receptores de Somatostatina/clasificaciónRESUMEN
OBJECTIVE: To study the expression of eIF4E, p-eIF4E (Ser 209) and Mcl-1 gene in the pathological scars and to investigate its role and its probable mechanism in the pathogenesis of abnormal scar. METHODS: Quantitative real-time PCR and Western Blot was performed to detect the expression and distribution of mRNA and protein of eIF4E and Mcl-1 in hypertrophic scar (10 cases), keloid (10 cases), normal scar (10 cases), and normal skin (10 cases). Western Blot was performed to detect the expression and distribution of protein of p-eIF4E in hypertrophic scar (10 cases), keloid (10 cases), normal scar (10 cases), and normal skin (10 cases). RESULTS: The expression of eIF4E mRNA and protein were 1.38 +/- 0.45, 1.23 +/- 0.23 in the normal skin (10 cases); 5.400 +/- 0.450, 5.460 +/- 0.460 in normal scar (10 cases); 0.597 +/- 0.060, 0.590 +/- 0.040 in hypertrophic scar (10 cases) and 0.694 +/- 0.066, 0.697 +/- 0.022 in keloid (10 cases). The expression of p-eIF4E protein in the normal skin (10 cases), normal scar (10 cases), hypertrophic scar (10 cases), and keloid (10 cases) were 0.202 +/- 0.037, 0.216 +/- 0.019, 0.426 +/- 0.026, 0.433 +/- 0.027. The expression of Mcl-1 mRNA and protein were 1.510 +/- 0.660, 1.400 +/- 0.530 in the normal skin (10 cases); 6.65 +/- 0.85, 7.23 +/- 1.53 in normal scar (10 cases); 0.589 +/- 0.059, 0.660 +/- 0.063 in hypertrophic scar (10 cases) and 0.870 +/- 0.118, 0.914 +/- 0.064 in the keloid (10 cases). The positive rate of mRNA and protein of eIF4E and Mcl-1 was not statistically different between the hypertrophic scar and keloid (P > 0.05), while they were all remarkably significant between normal scar and abnormal scar (P < 0.05). The phosphorylation of eIF4E in pathological scar was higher than that in control group. In pathological scar, mRNA and protein of eIF4E and Mcl-1 showed a strong positive correlation. CONCLUSIONS: The result indicates that the expression of eIF4E, p-eIF4E and Mcl-1 is increased in pathological scar. eIF4E plays an important role in pathological scar. Its activity is regulated by its phosphorylation. Therefore, eIF4E, p-eIF4E and Mcl-1 overexpression may play an important role in the proliferation of fibroblasts and in the pathogenesis of pathological scar.
Asunto(s)
Cicatriz/metabolismo , Factor 4E Eucariótico de Iniciación/metabolismo , Proteína 1 de la Secuencia de Leucemia de Células Mieloides/metabolismo , Adolescente , Adulto , Estudios de Casos y Controles , Factor 4E Eucariótico de Iniciación/genética , Femenino , Humanos , Queloide/metabolismo , Masculino , Proteína 1 de la Secuencia de Leucemia de Células Mieloides/genética , Fosforilación , ARN Mensajero/genética , Adulto JovenRESUMEN
OBJECTIVE: To study the expression of P57(kip2) and Maspin in the pathological scar and their possible role in the pathogenesis of abnormal scars. METHODS: Immunohistochemistry integrated image analysis and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of P57(kip2) and Maspin in hypertrophic scar, keloid, mature scar and normal skin. Statistics was used to analyze the datas. RESULTS: The expression of P57(kip2) protein was fixed to fibroblast intranuclear in abnormal scar, and the expression of P57(kip2) protein and P57(kip2) mRNA decreased (P < 0.05). The expression of Maspin protein was fixed to fibroblast cytoplasm and intranuclear in abnormal scar, and the expression of Maspin protein and Maspin mRNA decrease, compared with that in normal group (P < 0.05). There was positive correlation between P57(kip2) protein and Maspin protein expression (P < 0.01). CONCLUSIONS: The decreased expression of P57(kip2) and Maspin in abnormal scar shows that they are cicatrix-related genes. There is a positive relationship between the two genes. It may be one of the mechanisms of pathogenesis of abnormal scar. It makes effect through fibroblasts.
Asunto(s)
Cicatriz/metabolismo , Inhibidor p57 de las Quinasas Dependientes de la Ciclina/metabolismo , Serpinas/metabolismo , Cicatriz/patología , Cicatriz Hipertrófica/metabolismo , Cicatriz Hipertrófica/patología , Fibroblastos/metabolismo , HumanosRESUMEN
OBJECTIVE: To evaluate the application of temporal fascia flaps in the correction of severe depression deformities at lower eyelids. METHODS: Severe depression deformities at lower eyelid were corrected with temporal fascia flaps pedicled with superficial temporal artery in 9 cases. RESULTS: All flaps survived with good appearance. All patients were followed up for 6-12 months with good long-term results. The donor sites had no obvious scalp scar. CONCLUSIONS: Temporal fascia flap is an optimal choice for correction of the severe depression deformities at lower eyelid. It is easily performed with good result and less donor site morbidity.
Asunto(s)
Párpados/anomalías , Párpados/cirugía , Procedimientos de Cirugía Plástica/métodos , Adolescente , Adulto , Blefaroplastia/métodos , Femenino , Estudios de Seguimiento , Humanos , Masculino , Trasplante de Piel , Colgajos Quirúrgicos , Adulto JovenRESUMEN
OBJECTIVE: To introduce a new method for reconstruction of the whole ear lobe defect. METHODS: The free island skin flap supplied by superficial temporal vessel which was designed at the area anterior and superior to crus helicis. The flap was transferred through subcutaneous tunnel and self-folded to reconstruct the whole ear lobe defect. RESULTS: Since 1999, 6 cases were treated with no complication. The ear lobe shape and skin colour were very natural. CONCLUSIONS: The island skin flap supplied by superficial temporal vessel is very suitable for the whole ear lobe defect with good cosmetic results.
Asunto(s)
Oído Externo/cirugía , Procedimientos de Cirugía Plástica/métodos , Colgajos Quirúrgicos/irrigación sanguínea , Adulto , Oído Externo/patología , Femenino , Humanos , Masculino , Trasplante de Piel , Arterias Temporales/trasplanteRESUMEN
OBJECTIVE: To study the expression of Skp2 gene (s-phase kinase associated protein 2) in the pathological scars and its relationship with p27kip1 level and to investigate its role and its probable mechanism in the pathogenesis of abnormal scars. METHODS: Immunohistochemical technique was performed to detect the expression and distribution of Skp2 and p27kip1 protein in hypertrophic scar (42 cases), keloid (18 cases), normal scar (40 cases) and normal skin (50 cases), statistics was used to analyze the data. RESULTS: The positive rate of Skp2 and p27kip1 protein expression was not statistically different between the hypertrophic scar and keloid (P > 0.05), while they were all remarkably significant in comparison between normal scar and abnormal scar (P < 0.05). In pathological scar the protein of Skp2 and p27kip1 showed a strong inverse correlation (P < 0.01). CONCLUSION: The result indicated that the expression of Skp2 was increased and it may lead to decrease p27kip1 level in the hypertrophic scar and keloid, Skp2 overexpression might play an important role in the proliferation of fibroblasts and in the pathogenesis of pathological scar by adjusting the regulation of cyclins such as p27kip1.