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Objective: To investigate whether re-expression of protocadherin 10(PCDH10) induced by 5-aza-2'-deoxycytidine (5-Aza-CdR) could affect the invasion and migration of MDA-MB-231 cells, and to explore the possible mechanism. Methods: Human breast cancer cell line MDA-MB-231 was cultured in vitro. Control group and 5-Aza-CdR treatment group were set up. PCDH10 mRNA expression in MDA-MB-231 cell line was determined by reverse transcriptionpolymerase chain reaction (RT-PCR); Transwell chamber and wound healing assay were performed to measure the invasion and migration capacity of the cells, and protein expression of PCDH10, DNA methyltransferase(DNMT)3A, DNMT3B, nuclear factor(NF)-κB p65, matrix metalloproteinases(MMP)-2 and MMP-9 were detec Western blotting. Results: 5- Aza-CdR could reverse the methylation status of PCDH10 gene in MDA-MB-231 cells in a dose-dependent manner. Reexpression of PCDH10 significantly inhibited cell invasion and migration capacity in vitro. Western blottoing analysis revealed that the expression of DNMT3A, DNMT3B, NF-κB p65, MMP-2 and MMP-9 in MDA-MB-231 cells were downregulated after exposure to 5-Aza-CdR. Conclusion: Re-expression of PCDH10 significantly inhibits MDA-MB-231 invasion and migration capacity. The inhibitory effect is characterized that 5-Aza-CdR treatment down-regulates DNMT3A and DNMT3B levels, recovers the expression of anti-oncogene PCDH10, further blocks the activation of NF-κB p65, resultsing in a decrease in the secretion of MMP-2 and MMP-9.
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Objective: To investigate the expression of protocadherin 10 (PCDH10) in non-small cell lung cancer (NSCLC), and to explore the function and molecular mechanism of PCDH10 in NSCLC cells. Methods: The expression levels of PCDH10 and Ki67 mRNAs in NSCLC tissues and paracancerous normal tissues were detected by real-time fluorescent quantitative PCR. The correlations of PCDH10 and Ki67 mRNA expressions with the clinicopathological features and prognosis of NSCLC patients were analyzed. A549 cells were transfected with recombinant plasmid pcDNA3.1(+)-PCDH10 or empty vector pcDNA3.1(+) (as the control) by LipofectAMINE 2000. The proliferation, apoptosis and cell cycle were detected by IncuCyte S3 live-cell analysis system and FCM method, respectively. The migration and invasion abilities were detected by Transwell method. The expression levels of proliferating cell nuclear antigen (PCNA), Bcl-2, cyclin D1, cyclin E, cyclin-dependent kinase 4 (CDK4), E-cadherin and Slug mRNAs and proteins were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. Results: The expression level of PCDH10 mRNA in NSCLC tissues was lower than that of para-cancerous normal tissues (P < 0.001). The expression level of PCDH10 mRNA in NSCLC tissues was negatively correlated with T stage, lymph node metastasis, clinical stage and Ki67 mRNA expression (all P < 0.001), and positively correlated with prognosis of NSCLC patients (P < 0.001). PCDH10 over-expression inhibited the proliferation of A549 cells by blocking cell cycle at G1 phase and promoting apoptosis (all P < 0.05). PCDH10 significantly inhibited the invasion and migration of A549 cells (both P < 0.05). The expression levels of PCNA, cyclin D1, cyclin E, CDK4, Bcl-2 and Slug mRNAs and proteins were downregulated in A549 cells with PCDH10 over-expression (all P < 0.05), but the expression levels of E-cadherin mRNA and protein were up-regulated (both P < 0.05). Conclusion: PCDH10 is lowly expressed NSCLC tissues. PCDH10 over-expression can inhibit the proliferation and invasion of NSCLC cells.
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Background and purpose:Promoter methylation ofPCDH10, a gene encoding protocadherin 10, has been found to be correlated to poor prognosis in gastric cancer (GC) patients. However, the relationship between the expression of PCDH10 and prognosis in GC remained unknown. This study aimed to explore the relationship be-tween the expression of PCDH10 and clinicopathological features and prognosis of GC, and to identify biomarker for predictions of recurrence and survival of GC.Methods:mRNA expressions of PCDH10 in 115 pairs of GC tissues and adjacent normal tissues were detected by real-time lfuorescence quantitative polymerase chain reaction (RTFQ-PCR). The correlation between PCDH10 expression level and clinicopathological features and prognosis of GC was analyzed. Prediction models for 5-year recurrence and 5-year survival were established using logistic regression method.Results:Progression-free survival (PFS) and overall survival (OS) were signiifcantly prolonged in patients with PCDH10 low expression compared to patients without PCDH10 low expression (P=0.046 andP=0.033 respectively). PCDH10 low expression signiifcantly correlated with less lymph node metastasis (P=0.001) and earlier TNM staging (P=0.001), and was more common in female than in male (P=0.040). The mRNA expression of PCDH10 did not correlate with age, Lauren classiifcation, T stage, neural invasion or vascular invasion. Univariate Cox analysis showed Lauren classiifca-tion, T stage, N stage, M stage and PCDH10 expression signiifcantly correlated with PFS and OS. Logistic regression models for the prediction of 5-year recurrence or 5-year survival based on clinicopathological features included Lauren classiifcation, T stage, N stage and M stage as variables. Logistic regression models for the prediction of 5-year recur-rence or 5-year survival based on PCDH10 expression included Lauren classiifcation, T stage, M stage and PCDH10 expression level but not N stage as variables. The models based on PCDH10 expression had the same effciencies as models based on clinical parameters in predicting 5-year recurrence or 5-year survival for GC patients.Conclusion:PCDH10 low expression correlated with better prognosis, less lymph node metastasis and earlier TNM stage in GC patients. Low expression of PCDH10 may be a biomarker of better survival for GC patients. Logistic regression model based on PCDH10 mRNA expression may serve as a prediction model when patients have unknown lymph node metas-tasis status.