ABSTRACT
AIM:To observe the expression of calreticulin(CRT)in nasopharyngeal carcinoma tissues,ana-lyze the significance of clinical pathology and the influence on epithelial -mesencymal transition(EMT)of CNE2 cells. METHODS:The expression of calreticulin was detected by immunohistochemistry in 52 nasopharyngeal carcinoma and 57 nasopharyngeal benign tissues,and the significance of clinical pathology was evaluated.The calreticulin gene-specific small interfering RNA was constructed,and then was transfected into the NPC cell line CNE 2 using the cationic liposome meth-od.The effect of CRT on the morphological changes of the CNE 2 cells was observed under light microscope.The effect of CRT on the cell migration and invasion abilities of the CNE 2 cells was detected by Transwell migration and invasion assays. The expression of EMT-related proteins E-cadherin,vimentin,transforming growth factor(TGF)-βand matrix metallopro-teinase(MMP)-9 in the CNE2 cells was determined by Western blot.RESULTS:The positive expression rate of CRT in the benign lesion tissues was 19.29%(11/57),which was significantly increased in the nasopharyngeal carcinoma tissues as 82.69%(43/52).The expression rate of CRT was positively correlated with the stage of nasopharyngeal carcinoma and lymph node metastasis(P<0.05).Knockdown of CRT expression made the CNE 2 cells showing a spindle shape to a flat, cobblestone-like epithelial state change,arranged more compact,and the migration and invasion abilities were significantly decreased(P<0.05).Knockdown of CRT expression resulted in significant increase in the protein expression of E -cadhe-rin,and the decreases in the protein expression of vimentin, TGF-βand MMP-9 in the CNE2 cells(P<0.05).CON-CLUSION:Calreticulin expression in nasopharyngeal carcinoma is significantly higher and positively correlated with naso -pharyngeal carcinoma stage and lymph node metastasis.Calreticulin promotes cell migration and invasion of nasopharyngeal carcinoma CNE2 cells by inducing EMT.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the molecular mechanism of miR-124 suppressing the proliferation and invasion of gastric cancer cells.</p><p><b>METHODS</b>SPHK1 3'UTR-luciferase vector was constructed and luciferase reporter gene assay was employed to examine the effect of miR-124 on luciferase activity. Human gastric cancer MGC-803 cells were transfected with miR-124 mimics, and then Western blot was performed to detect the expression of SPHK1 protein.</p><p><b>RESULTS</b>Luciferase reporter vector system confirmed that SPHK1 was a target gene of miR-124. Western blot showed that the expression of SPHK1 protein was inhibited by miR-124. After transfection of miR-124 mimics or SPHK1 siRNA for 12 h, 24 h and 48 h, respectively, MTT assay showed that the A values of the three groups were significantly different (P < 0.05), and it was in a time-dependent manner. After transfection of miR-124 mimics or SPHK1 siRNA for 24 h, transwell invasion assay showed that the number of transmembrane cells was 54.6 ± 8.3 in the SPHK1 siRNA group and 47.8 ± 6.6 in the miR-124 mimics group, both were significantly lower than 100.6 ± 11.3 of the control group (P < 0.05), indicating that SPHK1 siRNA can slow down the invasion of MGC-803 cells.</p><p><b>CONCLUSION</b>miR-124 can suppress the cell proliferation and invasion by targeting SPHK1 in gastric carcinoma.</p>
Subject(s)
Humans , Cell Line, Tumor , Cell Proliferation , Genetic Vectors , Luciferases , Genetics , Metabolism , MicroRNAs , Genetics , Neoplasm Invasiveness , Phosphotransferases (Alcohol Group Acceptor) , Genetics , Metabolism , RNA, Small Interfering , Genetics , Recombinant Proteins , Genetics , Metabolism , Stomach Neoplasms , Metabolism , Pathology , TransfectionABSTRACT
Immunohistochemical technique was an essential tool of conventional diagnosis,therefore,the application of immunohisto- chemistry in reformation of pathology laboratory teaching would boost pathological experimental teaching standards to a higher level.