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@#Objective: To detect the expression of GRHL2 (grainyhead-like-2) in breast cancer tissues and to explore its correlation with clinicopathological characteristics and prognosis of breast cancer (BC) patients，aiming to find new therapeutic target for breast cancer. Method: A total of 88 pairs of BC tissues and corresponding para-cancerous tissues from patients with primary BC that treated and pathologically confirmed at the Second Department of General Surgery, Xinxiang Central Hospital from January 2010 to January 2017 were collected for this study. The expression of GRHL2 in BC tissues and para-cancerous tissues was examined with IHC, and the association between GRHL2 and clinicopathological characteristics of BC patients was analyzed. Moreover, the correlation between GRHL2 and prognosis of BC patients was investigated by analyzing TCGA clinic data for BC. Result: The expression of GRHL2 was significantly higher in BC tissues (75.00%) compared with para-cancerous tissues (36.36%) (P<0.01); Based on the results of GRHL2 expression in 114 cases of normal breast tissues and 1 097 cases of primary breast cancer tissues in TCGA database, the expression of GRHL2 in primary BC tissues was significantly higher than that in normal breast tissues (P<0.01). GRHL2 expression was associated with BC TNM stage,histological grade, HER2 status and lymphnode metastasis status (all P<0.05); TCGA database showed that the RFS of 1 979 BC patients with high GRHL2 expression was significantly shorter than that of the 1 972 cases of BC patients with low GRHL2 expression (HR=1.24, 95%CI:1.11-1.38, P<0.01); GRHL2 expression exerted no significant effect on RFS of TNBC patients or ER+ BC patients (TNBC: HR=1.30，95%CI: 0.89-1.88，P=0.170; ER+: HR=1.17, 95%CI:0.76-1.78， P=0.470); however, the RFS of HER2+ BC patients with high GRHL2 expression was significantly shorter than that of HER2+ BC patients with low GRHL2 expression (HR=1.72, 95%CI:1.11-2.68, P=0.015） . Conclusion：Expression level of GRHL2 was up-regulated in BC tissues, and was associated with BC TNM stage, histological grade, HER2 status and the lymphnode metastasis status. GRHL2 plays an important role in the generation and development of BC, indicating poor prognosis.
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Objective: To investigate the expression of grainyhead like 2 (GRHL 2) gene in human colorectal cancer (CRC) tissues and cells, and to explore the effects of GRHL 2 gene silence on the proliferation and cell cycle of CRC cells. Methods: The expression level of GRHL2 mRNA in CRC tissues and their para-cancerous normal tissues as well as CRC cells (HT29, SW480, SW1116, SW620, HCT116, Caco2 and LoVo cells) was detected by real-time fluorescent quantitative PCR. After the recombinant lentivirus carrying GRHL2-shRNA was infected into HT29 and HCT116 cells, the cell proliferation was determined by CCK-8 method and cloning formation assay, the cell cycle distribution was detected by FCM, and the expressions of cell cycle-related proteins (cyclin A, cyclin D1, p21 and p27) were determined by Western blotting. Furthermore, the subcutaneous transplantation models of CRC HT29 and HCT116 cells were established in nude mice, then the effect of GRHL 2 gene silencing on the growth of colorectal cancer xenografts was observed. Results: The expression level of GRHL2 mRNA in CRC tissues was higher than that in paracancerous normal tissues (2.8 times, P < 0.001). GRHL2 mRNA was highly expressed in most CRC cells (HT29, SW480, HCT116, Caco2 and LoVo cells). The expression levels of GRHL2 mRNA and protein in HT29 and HCT116 cells were down-regulated after transfection of GRHL2-shRNA (both P < 0.001). Silencing GRHL 2 gene significantly suppressed cell proliferation (both P < 0.05), produced the cell cycle arrest in G1 phase (both P < 0.01), down-regulated the expressions of cyclin A and cyclin D1 (P < 0.05, P < 0.01), and upregulated the expressions of p21 and p27 (P < 0.01, P < 0.05). Meanwhile, GRHL 2 gene silencing significantly promoted the growth of colorectal cancer xenografts in nude mice. Conclusion: GRHL2 maybe promote the proliferation of CRC cells by regulating the expression of cell cycle-related proteins.
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Objective:To investigate association between genetic polymorphism in the grainyhead-like 2 gene (GRHL2)and noise-induced hearing loss (NIHL)in the Chinese population.Methods:A matched case-control association study was employed,In which,3 790 workers exposed to continuous and steady-state occupational noise in a steel factory participated.The questionnaires were adopted to col-lect individual features and audiometry tests performed.In the sstudy,286 subjects were diagnosed as ca-ses,Which were each designated on the basis of the matched criterion,and 286 paired samples were se-lected finally.Noise intensity was measured according to the standards given in ‘Measurement of Noise in the Workplace’(Occupational Health Standard of the People’s Republic of China,GBZ /T1 89.8 -2007).Cumulative noise exposure (CNE)was calculated,according to monitoring data on A-weighed sound pressure level and employment time.Genomic DNA was obtained from peripheral blood samples using 2 mL DNA extraction Kit following the manufacturer’s protocol.Five single nucleotide polymor-phisms (SNPs)of GRHL2 were genotyped by multiplex SNP genotyping kit.The continuous variables and categorical variables were analyzed by t-test and chi-square test respectively.Multivariate Logistic re-gression was used to test the association between genetic frequency and disease status,with adjustments for the possible confounding variables.The haplotypes were established and their frequencies in the two groups were assessed by haploview and phase softwares.Results:All the five SNPs (rs373571 3, rs3824090,rs373571 4,rs373571 5 and rs61 1 41 9)were in Hardy-Weinberg equilibrium (HWE)(P >0.05).The subjects carrying rs373571 5 GG genotype had a higher NIHL risk than those carrying the GA genotype under the co-dominant model (OR =0.644,95% CI:0.442 -0.939,P =0.022)after ad-justment for height,blood pressure,drinking status and smoking status.After being stratified by CNE,in the CNE ≥ 98 dB (A)group,rs373571 5 polymorphism was associated with the NIHL under the co-dominant model (OR =0.509,95% CI:0.281 -0.923,P =0.026)after adjustment for height,blood pressure,drinking status and smoking status as well.However,no statistical significant difference was found in variant genotypes of the other SNPs between the case and control subjects.Four-locus (rs373571 3,rs3824090,rs373571 4 and rs373571 5)haplotypes were constructed,and no risk or protec-tive haplotypes was identified.Conclusion:It is suggested that GRHL2 polymorphisms may be associated with development of NIHL.