ABSTRACT
Objective To investigate the expression of LncRNA LINC00857 in pancreatic cells and the effect of lncRNA LINC00857 down-regulation on proliferation, migration and apoptosis of pancreatic cancer PANC-1 cells and possible mechanism. Methods The lentiviral vector GV112 was constructed and infected PANC-1 cells to obtain experimental group, while the blank plasmid was transfected as a negative control group and the cells without intervention were taken as normal control group. CCK-8 assay, Transwell assay, scratch test, flow cytometry, Western blot were used to detect the effect of LINC00857 down-regulation on cell proliferation, migration, apoptosis, cell cycle and EMT-related proteins. Results LINC00857 expression in pancreatic cancer cells were significantly higher than that in normal pancreatic epithelial cells (P < 0.001). Knockdown of LINC00857 could significantly inhibit the proliferation and migration of PANC-1 cells (P < 0.0001); compared with the negative control group, the apoptosis rates of cells in the experimental groups were significantly increased (P < 0.05), the number of cells in G0/G1 phase increased (P < 0.01), the number of cells in S phase decreased (P < 0.05), and the cells were blocked in the G1 phase, E-cadherin expression was significantly up-regulated (P < 0.05) and N-cadherin and Vimentin expression were significantly down-regulated (N-cadherin: P < 0.01, Vimentin: P < 0.05). Conclusion LINC00857 can promote proliferation and migration of PANC-1 cells and inhibit its apoptosis by regulating G1/S phase transition and EMT signaling pathway.
ABSTRACT
Secreted frizzled-related protein (SFRP) can inhibit the expression of Wnt signaling pathway through Frizzled protein.The silencing of SFRP gene promoter methylation is associated with the occurrence and metastasis of many cancers such as colorectal cancer,gastric cancer,liver carcinoma,lung cancer and ovarian cancer.Several studies have found that SFRP gene has latent clinical value,which is expected to become the novel target for the gene diagnosis and treatment of cancer.
ABSTRACT
Objective To investigate the correlation between invasion ability and cytoskeleton remodeling of hepatocarcinoma cell by atomic force microscopy (AFM) and to explore mechanical properties during genesis,development and invasion of hepatocellular carcinoma (HCC).Methods Four HCC cell lines (MHCC-97H,MHCC-97L,SMMC-7721 ,Huh-7 )with different invasive ability were studied.Mechanical parameter (Young′s modulus)was measured by AFM.The pattern of cytoskeleton remodeling of HCC cell lines with different invasive ability was detected by immunofluorescent staining. The difference of cell invasive ability was tested by cell scratch experiment in other to verify mechanical data.Kruskal-Wallis H test was used to compare the differences between groups.Results The results of AFM indicated that Young′s modulus of cytoplasma area and nucleus area decreased gradually as cell invasion ability increased (χ2 =472.78,622.43,both P <0.01).According to invasive ability from low to high,Young′s modulus of cell cytoplasm area of Huh-7,SMMC-7721 ,MHCC-97L and MHCC-97H were 1 602.43 (845 .48,3 317.25)Pa,1 055 .28 (367.48,2 280.77)Pa,1 026.78 (369.20,2 019.96)Pa and 503.12 (366.11 ,700.31)Pa,respectively.Young′s modulus of cell nucleus area of Huh-7,SMMC-7721 ,MHCC-97L and MHCC-97H were 2 823.98 (1 262.78,4 440.07 )Pa,1 313.43 (590.71 , 2 678.62)Pa,1 285 .17 (583.29,1 961 .19)Pa and 655 .57 (441 .29,943.39)Pa,respectively.The results of immunofluorescent staining showed that the stronger the cell invasive ability,the worse cytoskeletal integrity and more irregular cell microfilament distribution. In cell scratch assay, the migration rate of MHCC-97H was 46.67% in 24 h and 86.47% in 48 h,that of MHCC-97L was 45 .70%in 24 h and 82.86% in 48 h,that of SMMC-7721 was 39.41 % in 24 h and 79.85 % in 48 h and that of Huh-7 was 34.60% in 24 h and 72.09% in 48 h,which showed that the cell migration orderly in creased as the cell invasion ability increased.Conclusions It seemed that HCC with higher invasive ability had lower Young′s modulus,softer cell,stronger deformability,worse cytoskeleton integrity and more irregular cell structure,and vice versa.