Objective To study the cellular senescence and molecular mechanism of olaparib in MCF-7 breast cancer cells.Methods The effects of olaparib on the proliferation and migration of MCF-7 cells were detected dynamically by real-time cell analysis(RTCA)technology.The effects of olaparib on the Senescence was detected by using the senescence-associated β-galactosidase(SA-β-gal).Quantitative polymerase chain reaction was used to analyze the effects of olaparib on the expression levels of genes encoding the senescence-associated factors p16,p21,C/EBP homologous protein,interleukin(IL)-6,IL-8,plasminogen activator inhibitor 1,phosphatase and tensin homolog deleted on chromosome 10,p27,retinoblastoma gene,Ki67,and E2F1.The effects of olaparib on the expression levels of the senescence-associated proteins p21,γH2AX,pRB,cyclin D1,insulin-like growth factor binding protein 3,and Ki67 were analyzed by Western Blot.Results Olaparib inhibited the proliferation and migration and induced the senescence of MCF-7 cells.Long-term(96 h)treatment with olaparib significantly up-regulated the gene expression levels of p16,p21,p27,C/EBP homologous protein,IL-6,IL-8,plasminogen activator inhibitor 1,phosphatase and tensin homolog deleted on chromosome 10,and retinoblastoma protein(P＜0.01)and significantly down-regulated the gene expression levels of Ki67 and E2F1(P＜0.01)in MCF-7 cells.Olaparib significantly increased protein expression levels of p21,γH2AX,and insulin-like growth factor binding protein 3 in MCF-7 cells(P＜0.01,P＜0.01,P＜0.05)and significantly decreased cyclin D1,pRB,and Ki67 levels(P＜0.05,P＜0.01,P＜0.05).Conclusions Olaparib can inhibit proliferation and migration and induce senescence in MCF-7 breast cancer cells.

Objective:To investigate the effects of low-dose radiation(LDR)combined with human telomerase reverse transcriptase C-terminal polypeptide 27(hTERTC27)overexpression on the proliferation and apoptosis of lung cancer A549 cells,and to observe the in vivo antitumor effects of LDR combined with C27.Methods:The pEgr-hTERTC27 plasmid was transfected into human non-small cell lung cancer(NSCLC)A549 cells and mouse Lewis lung carcinoma(LLC)cells.Cells stably expressing C27(A549-C27 and LLC-C27)were screened by G418 selection.The cells were divided into six groups:conventional radiation(CONV-RT)group(A549-Con),LDR group(A549-Low),C27 group(A549-C27),CONV-RT combined with C27 group(A549-C27-Con),LDR combined with C27 group(A549-C27-Low),and control group(A549-Mock).The irradiation dose in LDR group was only 36%of CONV-RT group.MTT assay was used to detect cell proliferation,and flow cytometry was used to measure cell apoptosis.LLC cell-transplanted tumor model in mice was established through subcutaneous implantation,and the animal experiment groups were similar with cell experiments.Tumor growth,volume,and mass were recorded in each group.Muscle infiltration near the transplanted tumors was observed using H-E staining.Results:Compared with A549-Mock group,the proliferative activity of A549-Con and A549-Low group was significantly decreased(all P<0.01).Furthermore,the proliferation activity of A549-C27-Con and A549-C27-Low cells was significantly lower than that of A549-C27 cells(all P<0.01).Compared with A549-Mock,the apoptosis rates were significantly higher in A549-Con and A549-Low groups(P<0.01);however,no significant difference in apoptosis rates were observed among A549-C27,A549-C27-Con and A549-C27-Low groups(all P>0.05).In tumor-bearing mice,both CONV-RT and LDR significantly reduced tumor mass compared with unirradiated mice(all P<0.01).In addition,the tumor mass and local infiltration were significantly reduced in both LLC-C27-Low and LLC-C27-Con groups.Conclusion:LDR combined with C27 achieves similar antitumor effects to CONV-RT alone while reducing the total radiation dose in NSCLC.Moreover,LDR and C27 peptide synergize in their anti-tumor effects,with their combination reducing local tumor infiltration in transplanted tumor models.

Objective: To explore the genetic basis for a family with non-syndromic autosomal recessive deafness. Methods: The proband and her parents were subjected to physical and audiological examinations. With genomic DNA extracted from peripheral blood samples, next-generation sequencing was carried out using a panel for deafness genes. Suspected mutation was validated by Sanger sequencing and qPCR analysis of her parents. Results: The proband presented bilateral severe sensorineural hearing loss at three days after birth. Her auditory threshold was 110-120 dBnHL but with absence of vestibular and retinal symptoms. Her brother also had deafness but her parents were normal. No abnormality was found upon physical examination of her family members, while audiological examination showed no middle ear or retrocochlear diseases. Next-generation sequencing identified compound heterozygous mutations of the MYO7A gene, including a previously known c. 462C>A (p. Cys154Ter) and a novel EX43_46 Del, which were respectively derived from her mother and father. Conclusion The compound heterozygous mutations of the MYO7A gene probably underlie the disease in this family. Our findings has enriched the mutation spectrum for non-syndromic autosomal recessive deafness 2.

OBJECTIVE: To explore the clinical and genetic features of a patient suspected with Juvenile Parkinson's syndrome (JP). METHODS: Clinical features of the patient were analyzed. Genomic DNA of the patient and his parents was extracted from peripheral blood samples and sequenced by exome capture sequencing. The nature and impact of detected mutations were predicted and validated. RESULTS: The patient displayed typical features including resting tremor, bradykinesia, rigidity, but with excellent response to low dose levodopa. DNA sequencing showed that she has carried compound heterozygous mutations of the Parkin gene, namely c.1381dupC and c.619-1G>C, which were respectively inherited from his mother and father. Neither mutation was reported previously. Bioinformatic analysis predicted that both mutations are pathogenic. CONCLUSION The patient has JP caused by mutations of the Parkin gene. Exome capture sequencing is an accurate and efficient method for genetic diagnosis of such disease.
Adolescent ; Base Sequence ; Female ; Humans ; Mutation ; Parkinson Disease ; Ubiquitin-Protein Ligases ; Whole Exome Sequencing

OBJECTIVE: To explore the genetic basis for a family with non-syndromic autosomal recessive deafness. METHODS: The proband and her parents were subjected to physical and audiological examinations. With genomic DNA extracted from peripheral blood samples, next-generation sequencing was carried out using a panel for deafness genes. Suspected mutation was validated by Sanger sequencing and qPCR analysis of her parents. RESULTS: The proband presented bilateral severe sensorineural hearing loss at three days after birth. Her auditory threshold was 110-120 dBnHL but with absence of vestibular and retinal symptoms. Her brother also had deafness but her parents were normal. No abnormality was found upon physical examination of her family members, while audiological examination showed no middle ear or retrocochlear diseases. Next-generation sequencing identified compound heterozygous mutations of the MYO7A gene, including a previously known c.462C>A (p. Cys154Ter) and a novel EX43_46 Del, which were respectively derived from her mother and father. CONCLUSION The compound heterozygous mutations of the MYO7A gene probably underlie the disease in this family. Our findings has enriched the mutation spectrum for non-syndromic autosomal recessive deafness 2.
Female ; Hearing Loss, Sensorineural ; genetics ; High-Throughput Nucleotide Sequencing ; Humans ; Male ; Mutation ; Myosins ; genetics ; Pedigree