Expression of nestin, transcription factor SOX3 and homeobox protein DLX-4 in the facial tissue of children with cleft lip and palate / Expresión de nestina, factor de transcripción SOX3 y proteína homeobox DLX-4 en el tejido facial de niños con fisura labiopalatina

The failure of fusion of nasal and maxillary processes results in cleft lip and palate (CLP), which is one of the most common birth defects. The morphopathogenesis of this pathology is multifactorial and still largely unclear. The aim of this study was to evaluate the presence of nestin, transcriptor factor SOX3 (Sox3) and homeobox protein DLX-4 (Dlx-4) in complete unilateral (CU) and complete bilateral (CB) CLP affected facial tissue. Oral mucosa tissue samples were obtained from 17 CUCLP and 13 CBCLP patients during surgical cleft correction and 6 unaffected control subjects. Obtained tissue sections were stained with hematoxylin and eosin and by immunohistochemistry for nestin, Sox3 and Dlx-4. The intensity of staining was graded semiquantitatively. Nestin-positive structures were detected in all CUCLP and CBCLP patients' tissue samples, varying from moderate number of nestin-positive structures to numerous. Sox3 immunoreactivity was more prominent in epithelial cells in both patient groups with frequently patchy distribution. Mainly moderate number of Dlx-4-positive cells was observed in most of tissue samples. Statistically significant moderate positive correlation was found between nestin and Sox3 factors in CUCLP patient group (Spearman's rank correlation coefficient = .517, P = .034). Increase of nestinpositive structures suggests its role in the regulation of the remodeling of tissue in both CUCLP and CBCLP affected tissue. Dominance of Sox3 positivity in cleft affected epithelium indicates its possible role in (compensatory) formation of defective oral epithelium of CUCLP and CBCLP patients. The reduced expression of Dlx-4 implicates its limited regulatory role on the craniofacial development in CUCLP and CBCLP affected tissue.

La falla en la fusión de los procesos nasal y maxilar son causante de la fisura labiopalatina (FLP), que es uno de los defectos congénitos más comunes. La morfopatogenia de esta patología es multifactorial y aún poco clara. El objetivo de este estudio fue evaluar la presencia de nestina, el factor transcriptor SOX3 (Sox3) y la proteína homeobox DLX-4 (Dlx-4) en todo el tejido facial afectado por FLP bilateral unilateral (FU) y bilateral completa (FB). Se obtuvieron muestras de tejido de mucosa oral de 17 pacientes FUFLP y 13 FBFLP durante la corrección quirúrgica de la fisura y de 6 sujetos de control no afectados. Las secciones de tejido obtenidas se tiñeron con hematoxilina y eosina y mediante inmunohistoquímica para nestina, Sox3 y Dlx-4. La intensidad de la tinción fue graduada semicuantitativamente. Se detectaron estructuras positivas para nestina en todas las muestras de tejido de pacientes FUFLP y FBFLP, variando desde un número moderado a numerosas estructuras positivas para nestina. La inmunorreactividad de Sox3 fue más prominente en las células epiteliales en ambos grupos de pacientes con distribución frecuentemente irregular. Se observó un número principalmente moderado de células Dlx-4-positivas en la mayoría de las muestras de tejido. Se encontró una correlación positiva moderada estadísticamente significativa entre los factores de nestina y Sox3 en el grupo de pacientes FUFLP (coeficiente de correlación de rangos de Spearman = 0,517, P = 0,034). El aumento de estructuras positivas para nestina sugiere su papel en la regulación de la remodelación de tejido, tanto en tejido afectado por FUFLP como FBFLP. La dominancia de la positividad de Sox3 en el epitelio afectado de la fisura, indica su posible papel en la formación (compensatoria) del epitelio oral defectuoso de pacientes FUFLP y FBFLP. La expresión reducida de Dlx-4 implica su función reguladora limitada en el desarrollo craneofacial en tejido afectado por FUFLP y FBFLP.

The effect of DLX4 on the apoptosis of choriocarcinoma cells and its related mechanism / 肿瘤

Objective: To explore the effect of homeodomain protein DLX4 on apoptosis of choriocarcinoma cells and its related mechanism. Methods: RNA interference (RNAi) was used to knockdown DLX4 expression in JEG-3 cells. Then the inhibitory effect of DLX4 on apoptosis was determined by flow cytometry and the expression of apoptosis-related protein cleaved caspase-3 and caspase-8 and bax was detected by western blotting. Results: RNA interference targeted for DLX4 effectively and specifically inhibited DLX4 mRNA expression and decreased DLX4 protein level in JEG-3 cells. The apoptotic rate of JEG-3 cells was increased, the expression levels of cleaved both caspase-3 and caspase-8 were elevated but the bax protein expression did not change after knocking down DLX4. Conclusion: RNA interference silences DLX4 expression and induces apoptosis of JEG-3 cells. The mechanism for apoptosis was related with transcriptional regulation on caspase-3 and caspase-8.

The inhibitory effect of DLX4 RNAi on adhesion and invasion ability of choriocarcinoma cells / 中国癌症杂志

Background and purpose:Choriocarcinoma is a highly malignant tumor,which metastasis could occur at early stage,the molecular mechanism is still under investigation.DLX4 is a member of homeodomain transcriptional factors,and its abnormal expression has a close correlation with mammary cancer,colon caner and so on.Based on our previous study that DLX4 was highly expressed in choriocarcinoma JEG-3 cell lines,we used RNAi to knock down the expression of DLX4,and aimed to explore the effect of DLX4 on the adhesion,movement and invasion ability of choriocarcinoma cells.Methods:DLX4 siRNA and control siRNA plasmid were constructed and transfected into choriocarcinoma cell line JEG-3 cells with LipofectamineTM2000.JEG-3 cells that expressed high level of DLX4 siRNA and control siRNA plasmid were selected and maintained with G418.The effect of RNAi was detected by western blot,and then the adhesion,movement and invasion ability of JEG-3 cells were investigated by cell-ECM adhesion assay,Transwell chamber and Transwell-Matrigel model,respectively.Results:DLX4 RNAi inhibited DLX4 protein expression specifically and effectively.The adhesion rate of the cells in JEG-3 siDLX4 group was(30.6?4.2)%,which was significantly less than(57.2?4.0)% in JEG-3 without transfenction and(51.6?3.9)% in JEG-3 scDLX4 groups(P0.05).Conclusions:The knock-down of DLX4 by RNAi could prevent the adhesion and invasion ability of choriocarcinoma cells,which provides a new thought for further research in invasion and metastasis of choriocarcinoma.