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1.
Basic & Clinical Medicine ; (12): 1155-1161, 2015.
Article in Chinese | WPRIM | ID: wpr-479335

ABSTRACT

Objective To investigate the molecular mechanisms of PLCεin regulating the invasion and migration of human bladder cancer cells in vitro.Methods After cells treated with recombinant adenovirus , the migratory/in-vasive abilities of T24 cells were explored by wound healing and Transwell chamber cell migration and invasion as -say;RT-PCR was used to detect the mRNA levels of PLCε;The protein levels of PLCε,PKCα,PKCβ, TBX3 and E-cadherin were determined by Western blot;QRT-PCR was used to detect the mRNA levels of TBX3 and E-cad-herin.Results It was confirmed by digesting and sequencing that the recombinant adenovirus had been constructed successfully .The expression of PLCε mRNA and PLCε protein were both decreased after the infection of Ad-shPLCε.Wound healing and Transwell chamber cell migration/invasion assay showed that Ad-shPLCε treatment could inhibit the migratory and invasive activity of bladder cancer cells(P<0.05).The results of Western blot indicated that the expression of PKCα/βin membrane decreased ( P<0.05 ) , and phosphorylation level of PKCαand PKCβwas reduced .QRT-PCR and Western blot analysis demonstrated that the expression level of TBX 3 de-creased , but the expression level of E-cadherin increased .Conclusions PLCε shRNA can inhibit migratory and invasive ability of bladder cancer cells through PKCα/β/TBX3/E-cadherin pathway .

2.
Basic & Clinical Medicine ; (12): 208-212, 2015.
Article in Chinese | WPRIM | ID: wpr-480614

ABSTRACT

Objective To explore the impact of silencing PLCε1 gene on proliferation and cell cycle of esophageal carci-noma Eca109 cells.Methods Three plasmid expression vectors (PLCε11, PLCε12 and PLCε13) were constructed to si-lence PLCε1 gene.A negative control plasmid expression vector (HK) was constructed at the same time to serve as a control .The plasmid expression vectors were transfected into esophageal carcinoma Eca 109 cells by cations liposome . The plasmid expression vector with the best interference effect ( PLCε12 ) was chosen .The study included Eca 109 group , HK group and PLCε12 group .Cell viability of Eca 109 cells was evaluated by MTT assay .The cell cycles were detected by FCM .The mRNA expression of P16 and CyclinD1 gene was measured by RT-PCR.Results The cell vi-abilitys of Eca109 cells in PLCε12 group were 80.73%and 75.88%at 48 and 72 h after transfection , which were significantly lower than that of Eca 109 cells in HK group (P<0.001).The percentage of S phase Eca109 cells in PLCε12 group was lower than that of Eca 109 cells in HK group ( P <0.01 ) , the cell cycle of PLCε12 group Eca109 cells was arrested in G0/G1 phase.The P16 gene mRNA expression of PLCε12 group Eca109 cells was higher than that of HK group Eca 109 cells ( P<0.01 ) .Conclusions Silencing PLCε1 gene may up-regulate P16 gene mRNA expression and then arrest the cell cycle at G 0/G1 phase and so inhibit proliferation of Eca 109 cells.

3.
Chinese Pharmacological Bulletin ; (12): 1388-1391,1392, 2014.
Article in Chinese | WPRIM | ID: wpr-599555

ABSTRACT

Aim To investigate the effects of PLCε on the invasion and migration of human osteosarcoma cancer cells U2OS. Methods RNA interference ( RNAi) was used to inhibit PLCεexpression, and the proliferation of cancer cells was measured by CCK-8 assay. The migration of the cells was measured by scratch wound healing assay and migration chamber as-say. Gelatin zymography was performed to measure the MMP2 activities in U2OS cells. Results PLCε ex-pression was suppressed by siRNA. CCK-8 assay showed that PLCε had no effect on the proliferation of cancer cells. PLCε knockdown inhibited cell invasion and activities of MMP2 . Conclusion PLCε knock-down can inhibit the migration and invasion of human osteosarcoma cancer cells U2OS.

4.
Chinese Journal of Clinical Oncology ; (24): 1437-1441, 2014.
Article in Chinese | WPRIM | ID: wpr-458283

ABSTRACT

Objective: To explore the association of PLCε1 gene rs2274223 A/G single nucleotide polymorphism (SNP) and rs11599672 T/G SNP with susceptibility to esophageal squamous cell carcinoma (ESCC) in a population of Ci County high-incidence region in Hebei Province. Methods:The genotypes of PLCε1 gene rs2274223 A/G SNP and rs11599672 T/G SNP were determined by polymerase chain reaction–ligase detection reaction method in 527 ESCC patients and 527 healthy controls. Results:The frequency of positive family history of upper gastrointestinal cancer UGIC in ESCC patients was 48.6%, which is significantly higher than that in the healthy controls (39.3%) (χ2=9.25, P=0.002). The AA, AG, and GG genotype frequencies of PLCε1 gene rs2274223 A/G SNP were 48.0%, 43.9%, 8.1%in the ESCC patients and 57.1%, 37.5%, 5.4%in the healthy controls, respectively. Compared with AA genotype, AG, GG, and AG/GG genotypes enhanced the risk of ESCC. The age, sex, smoking status, and UGIC family history-adjusted OR were 1.41 (95%CI=1.09-1.83), 1.71 (95%CI=1.03-2.86), and 1.45 (95%CI=1.13-1.85), respectively. No significant difference was observed in the frequency of the genotype and allele of PLCε1 gene rs11599672 T/G SNP between the ESCC cases and the controls (P>0.05). PLCε1 gene rs2274223 A/G SNP and rs11599672 T/G SNP were combined for analysis using a 2LD software. Results showed that no linkage disequilibrium exists between these two SNPs (D'=0.11). Compared with the most frequent AT haplotype, the GT haplotype sig-nificantly increased the risk of ESCC (OR=1.36, 95%CI=1.08-1.71). Conclusion:PLCε1 gene rs2274223 A/G SNP might serve as a marker predicting genetic susceptibility to ESCC of the population from Ci County. The subjects with UGIC family history and the AG-or GG-genotype carriers had higher risk of ESCC and should receive periodic upper gastrointestinal fiber tests for early detection and treatment of ESCC.

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