Clinical and genetic study of a family with cerebro-oculo-facio-skeletal syndrome / 中华实用儿科临床杂志

Objective To analyze the clinical features and pathogenic gene mutation in a Chinese family with cerebro-oculo-facio-skeletal(COFS)syndrome,in order to summarize the relationship between phenotype and genotype.Methods The clinical data of the proband and his family members were collected.Genomic DNA from the proband and his parents were extracted by using standard procedures from the peripheral blood leukocytes.Next-generation sequencing was used to detect gene mutation in the patient with COFS syndrome.Sanger sequencing was applied to confirm the results.Results The proband,male,1 year and 3 months old,presented with microcephaly nystagmus,large ears,prominent nose,high arched palate,overhanging upper lip,micrognathia,widely set nipples,flexion contractures(especially involving the elbows and knees),failure to thrive,developmental retardation and feeding difficulty.His parents were normal phenotype.Two different heterozygous mutations c.1843G>T(P.G615W)and c.1996 C> T(P.R666W)were identified in the ERCC2 gene.The proband's father had the heterozygous mutation c.1843G>T(P.G615W)and his mother had the heterozygous mutation c.1996 C> T(P.R666W).Meanwhile,this heterozygous mutation c.1996 C> T(P.R666W)had been reported as a pathogenic gene mutation.Conclusions COFS syndrome is characterized by microcephaly,prominent nose,arthrogryposis and severe developmental delay.This is the first report on COFS syndrome patient in the mainland of China.The pathogenic gene mutations and gene status were identified through genetic studies.The result has laid the foundation for accurate genetic counseling and further prenatal diagnosis.

Association between ERCC2/XPD Polymorphisms and UVC-induced DNA Damage Using Transfected Cells Model / 中国医科大学学报

Objective To explore the function of ERCC2/XPD polymorphisms in the repair of DNA damage induced by UVC. Methods Plas?mids stably expressing ERCC2/XPD rs13181 AA(Lys751)and ERCC2/XPD rs13181 CC(Gln751)were transfected into Chinese hamster ovary cells,and the stable ERCC2 transfected cell lines were obtained. MTT assay was used to compare the inhibitory rates of the transfected cells treated with UVC at different irradiation intensity. The DNA damage repair ability of the transfected cells treated with UVC for 1,3,6 and 24 h was detected by modified comet assay. Results Compared with UV5ERCC2(CC),UV5ERCC2(CC) was more sensitive to UVC with decreased cell viability. DNA damage level of UV5ERCC2(CC) cells was more serious than UV5ERCC2(CC). Conclusion DNA repair capacity of ERCC2/XPD rs13181A allelic is lower than its wild?type,suggesting that ERCC2/XPDpolymorphisms play a critical role in UVC?induced DNA damage repair.

Application of ERCC2 gene polymorphism detection in molecular pathological diagnosis of breast cancer / 临床与实验病理学杂志

Purpose To evaluate the application value of ERCC2 gene polymorphism ( rs3916840 C/T, rs1799793 G/A and rs238416 G/A) detection in molecular pathological diagnosis of breast cancer. Methods The polymorphisms of ERCC2 ( rs3916840, rs1799793 and rs238416) were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay in 101 patients with breast cancer and in 101 cancer-free controls. Results Analysis of the data showed a 0. 287-fold increased risk of breast cancer due to the deletion of genotype GA at rs238416 (P0. 05). Furthermore, Heterozygous genotype of rs3916840 was significantly associated with tumor size (P=0. 049), heterozygous genotype of rs1799793 was significantly associated with PR sta-tus and triple negative breast cancer (P=0. 037). Remarkably, the genotype frequency of GA in p53-positive patients was lower than that in p53-negiative patients (P=0. 026). Conclusions These results indicate that the polymorphism of rs238416 of ERCC2 is sig-nificantly associated with breast cancer risk. Tumor size, PR status, triple negative breast cancer, and p53 protein expression are asso-ciated with polymorphisms of ERCC2 (rs3916840, rs1799793 and rs238416) respectively. ERCC2 gene polymorphism detection is useful for the early diagnosis and prognosis evaluation of breast cancer.

Relationship between the expressions of ERCC1 and ERCC2 in peripheral venous blood and cancer tissues of esophageal squamous cell carcinoma patients / 肿瘤

Objective:To investigate the feasibility of detecting excision repair cross-complementing 1(ERCC1)and ERCC2 in peripheral venous blood instead of cancer tissues from esophageal squamous cell carcinoma patients. Methods:The expressions of ERCC1 and ERCC2 mRNA were detected by using RT-PCR in 39 cases of peripheral venous blood samples, esophageal squamous cell carcinoma tissues, and adjacent normal tissues. ELISA was used to determine the levels of ERCC1 and ERCC2 proteins in serum. The periphe-ral blood from 10 healthy volunteers was used as control. Results:Expression levels of ERCC1 and ERCC2 mRNA and protein were significantly higher in peripheral blood from healthy control than those in esophageal carcinoma patients (P<0.05). There was a positive correlation between the expression of ERCC1 and ERCC2 mRNA in peripheral blood and cancer tissues (P<0.01). Conclusion:The expression levels of ERCC1 and ERCC2 mRNA in peripheral blood can indirectly reflect their expression levels in human esophageal squamous cell carcinoma tissues.

ERCC2 expression in siRNA interference of esophageal cancer cells and correlation with paclitaxel sensitivity / 中国癌症杂志

Background and purpose: ERCC2 gene silence by siRNA interference was observed in esophageal cancer cell line and it could change cell sensitivity to taxol. This study was to investigate the biological mechanism of paclitaxel-resistance in esophageal cancer cells. Methods: ERCC2 targeting siRNA (si-ERCC2) has been synthesized. The constructor were transfected into ERCC2 cell lines high-KYSE150(bigh expression of ERCC2) through lipofectamine. RT-PCR and flow cytometry (FCM) were used to detect the ERCC2 mRNA and protein expression levels. MTr assay was used to estimate the paclitaxel sensitivity of the cells. Results: In si-ERCC2 group, ERCC2 could not be detected and ERCC2 protein expression were reduced by 31.2%, 51.6% and 60.0%, respectively, 24,48,72 b after transfection. Paclitaxel IC_(50) value for si-ERCC2 group was 6.32±0.87 μg/mL, lower than the control group(49%). Conclusion: siRNA could successfully silence the target gene ERCC2 at the level of transcription and translation of the gene, the reduction of ERCC2 expression may reverse taxol resistance of the cells.

Effect of silencing ERCC2 expression by siRNA interference on sensitivity of esophageal cancer cells to paclitaxel / 肿瘤

Objective:To silence ERCC2 (excision repair cross-complementing rodent repair deficiency,complementatin group 2,ERCC2) expression in esophageal cancer KYSE150 cells by small interfering RNA (siRNA) and observe the altered sensitivity of KYSE150 cells to paclitaxel (PTX) and elucidate the mechanism underlying the reversion of the PTX resistance of KYSE150 cells. Methods:ERCC2-targeted siRNA was synthesized in vitro and transiently transfected into ERCC2 overexpressing KYSE150 cells via Lipofectamine mediation. The mRNA and protein expression levels of ERCC2 were determined by using RT-PCR and FCM method, respectively. The sensitivity of KYSE150 cells to PTX was measured by MTT assay before and after siRNA transfection. Results:RT-PCR results suggested that the specific bands of ERCC2 mRNA were not detected in si-ERCC2 group at 24, 48 and 72 h post transfection. FCM results indicated that the expression levels of ERCC2 protein gradually decreased by 31.2%, 51.6% and 60.0% at 24, 48 and 72 h respectively(P<0.01). The IC_(50) value of PTX for ERCC2-silencing KYSE150 cells was (6.32±0.87) μg/mL, lower than that for control cells (P<0.01). Conclusion:siRNA successfully silenced the expressions of target gene ERCC2 at both the transcription and translation levels. Silencing ERCC2 expression partly reversed the resistance of KYSE150 cells to PTX.

Polymorphisms in ERCC1 and ERCC2/XPD and Survival in Non-Small-Cell Lung Cancer Patients Treated with Cisplatin Based Chemotherapy

PURPOSE: The expressions of low levels of ERCC1 (excision repair crosscomplementation group 1) and ERCC2/XPD (excision repair cross- complementation group 2) have been studied in order to find a potential marker for predicting the prognosis or treatment response in cancer patients. However, polymorphisms in these genes have been rarely evaluated in terms of predicting the survival of cancer patients. MATERIALS AND METHODS: We investigated whether these polymorphisms had an effect on the response to chemotherapy and on the survival in 109 patients, with non-small-cell lung cancer, treated with cisplatin plus gemcitabine, paclitaxel or docetaxel. The polymorphisms of ERCC1 Asn118Asn (C->T), ERCC2 Lys751Gln and Asp312Asn were evaluated using a SNaPshot kit. RESULTS: The treatment responses showed no statistically significant differences according to the polymorphisms of ERCC1 Asn118Asn, ERCC2 Lys751Gln or Asp312Asn. The median survival time was 376 days (95% CI, 291~488). The overall survival rate showed no significant difference according to age, sex, chemotherapy regimen, clinical stage or sequential radiation therapy. The polymorphisms of ERCC2 Lys751Gln and Asp312Asn did not affect the survival of the patients (p=0.4711 and 0.4542, respectively). The polymorphism of ERCC1 Asn118Asn, chemotherapy response, performance status and body weight loss had effect on the overall survival of the patients (p=0.0001, 0.0001, 0.0176 and 0.0082 respectively). As for survival rate, according to the polymorphism in ERCC1 Asn118Asn, the median survival time in those patients showing the wild genotype (C/C) was 480 days (95% CI, 333~544), which was statistically significant compared with the 281 days for the patients with the variant genotype (T/T, C/T) (hazard ratio 3.497) (95% CI, 214~376). CONCLUSION: It is suggested that the presence of the wild genotype in ERCC1 Asn118Asn, in non-small-cell lung cancer patients treated with cisplatin based chemotherapy, was a surrogate marker for predicting a better survival.