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Objective:To investigate the effect of miR-34a on the migration and invasion of prostatic cell line through MSR1.Methods:Western blot was used to detect the expression of MSR1 in prostate cancer cell lines.qPCR was used to detect the ex-pression of miR-34a in prostate cancer cell lines.The correlation between miR-34a and MSR1 was examined by double luciferase as-say.Transwell invasion assay was used to detect the effect of miR-34a and MSR1 on the invasion ability of prostate cancer cells.The effects of miR-34a and MSR1 on the fine migration ability of prostate cancer were examined by scratch healing.Results:The expression level of MSR1 in PC3 cells was relatively high by Western blot.The expression level of miR-34a in PC3 cells was relatively low by qPCR.Double luciferase assay showed a direct binding site between miR-34a and MSR1,MiR-34a could regulate the expression level of MSR1,Scaffold Healing Test and Transwell Invasion Test were used to detect the migration and invasion of PC3 cells after overexpressing miR-34a.Overexpression of MSR1 could reverse the inhibitory effect of miR-34a on the migration and invasion of PC3 cells.Conclusion:miR-34a can regulate the migration and invasion of prostate cancer cells through MSR1 protein.
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Objective To investigate the effects of isoliquiritigenin on the invasive ability of human gastric carcinoma SGC7901 cells, and its molecular mechanisms thereof. Methods The logarithmic phase human gastric carcinoma SGC7901 cells were divided into control group (normal cell culture fluid) and isoliquiritigenin group (isoliquiritigenin solu?ble in cell culture fluid, the concentrations were 10, 25, 50 and 100 μmol/L respectively). Each group had four repeated holes. The proliferation of SGC7901 cells were detected with MTT assay after 24 h, 48 h and 72 h of culture. The experimen?tal drug concentration and action time were researched for the subsequent experiments. The in vitro invasion abilities of SGC7901 cells were assessed with Transwell test. The expression levels of MMP9, Akt and P-Akt were detected by Western blot assay. Results The proliferation of SGC7901 cells were inhibited by 10μmol/L isoliquiritigenin, which can be signifi?cantly inhibited by 25, 50 and 100μmol/L isoliquiritigenin in a concentration-dependent and time-dependent manner. The half inhibitory concentrations (IC50) of 24, 48 and 72 h were 52.48, 44.49 and 32.50μmol/L, respectively. Therefore, the 25, 50 and 100μmol/L isoliquiritigenin were selected as the subsequent experimental drug concentration, and 24 h was used as the action time. Compared with the control group (209.75±9.29), the membrane cell number of 25μmol/L (138.50±10.15), 50μmol/L (89.50 ± 16.56) and 100μmol/L (45.00 ± 8.08) decreased gradually (F=267.948,P<0.05). There was no signifi?cant difference in the expression level of Akt protein between four groups (F=1.492). The expression levels of P-Akt and MMP9 were gradually decreased with the increase of the isoliquirigenin concentration (F=359.219 and 431.324,P<0.05). Conclusion Isoliquiritigenin can obviously inhibit invasion ability of SGC7901 cells, which may be related to the down reg?ulation of the signal transduction pathway protein PI3K/Akt and the down steam protein MMP9.
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To construct expression vectors of small hairpin RNA aimed at N-acetylglucosaminyltransferase Ⅴ(GnT-Ⅴ) gene, and to investigate effects of GnT-Ⅴ shRNA on proliferation, adhesion, migration and invasion of LoVo cell line. siRNAs were designed according to the coding sequence of GnT-Ⅴ gene, shRNA expression vectors were constructed and transfected into LoVo cell line, cell lines which stably expressed low level of GnT-Ⅴ were established by G418 screening. The mRNA and protein expression of GnT-Ⅴ were measured by semi- quantitative reverse transcription polymerase chain reaction(RT-PCR) and Western blot analysis, respectively. The effects of pGPU6/GFP/Neo GnT-Ⅴ shRNA vectors on proliferation, adhesion, migration and invasion of LoVo cell line were evaluated by CCK-8 assay, heterogenous adhesion, wound closure assay, chemotactic migration and cell invasive experiment, respectively. GnT- Ⅴ shRNA expression plasmid was constructed successfully and pGPU6/GFP/Neo GnT-Ⅴ shRNA down-regulated expression of GnT-Ⅴ dramatically in LoVo cell. Expression of LoVo GnT-Ⅴ/1564 and LoVo GnT-Ⅴ/2224 dereased by 82%, 71.5% respectively at mRNA level, and 68%, 56% respectively at protein level. The more effective interfered cell line, LoVo GnT-Ⅴ/1564, was chosen to do further experiment. CCK-8 assay showed proliferation of LoVo GnT- Ⅴ/1564 was suppressed obviously, compared to proliferation of negative control group cell (P
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Objective To study the effect of down-regulation of E-cadherin on the invasive ability of tumor cells. Methods Human pancreatic carcinoma cell lines JHP-1, PANC-1, and MIA PaCa-2 were selected. The immunocytochemistry, Western blot and invasive-MTT assay were used to observe the expression and the contents of E-cadherin in the tumor cells. Results Immunocytochemistry showed that E-cadherin expression was on cell membrane. The expressions of E-cadherin were preserved in JHP-1, reduced in PANC-1, and showed negative in MIA PaCa-2. Western blot analysis showed that the protein content was the highest in JHP-1 and lowest in MIA PaCa-2. According to invasive MTT assay, the invasive ability of MIA PaCa-2 was the strongest, followed by PANC-1 and JHP-1 (P