ABSTRACT
SWI1 is a member of a new class of tumor DNA-binding proteins named as the AT-rich interaction domain family (ARID), and considered to bind with AT base pairs specifically. Genomic and functional data support ARID1A as a tumor suppressor because ARID1A/BAF250a (SWI1) subunit of the SWI/SNF chromatin-remodeling complex has emerged as recurrently mutated in a broad array of tumor types. But the crystal structure of SWI1 has not been solved as yet. Using docking and molecular dynamics, we predicted the DNA interaction pattern of human SWI1 ARID and made comparisons with the other two representative ARID family members, human Mrf-2 ARID and Drosophila Dri ARID. Dynamic results revealed that the N-terminal and loop L1 of SWI1 ARID bound with the DNA major groove, while the loop L2 and helix H6 bound with the minor groove. Moreover, it was found that SWI1 ARID bound with DNA apparently in a sequence-nonspecific manner. It was concluded that SWI1 ARID can form stable complex with sequence-nonspecific DNA segment comparing to Mrf-2 ARID/DNA and Dri ARID/DNA sequence-specific complexes.
Subject(s)
Humans , Binding Sites , DNA , Chemistry , Metabolism , DNA-Binding Proteins , Chemistry , Metabolism , Drosophila Proteins , Chemistry , Homeodomain Proteins , Chemistry , Models, Molecular , Molecular Docking Simulation , Molecular Dynamics Simulation , Nuclear Proteins , Chemistry , Protein Structure, Tertiary , Transcription Factors , Chemistry , MetabolismABSTRACT
This study was purposed to investigate the relationship between expression of the FANCG gene and adult sporadic acute myeloid leukemia (AML), real-time PCR with SYBR Green I technique was used for detecting FANCG gene expression level in bone marrow mononuclear cells of 54 newly diagnosed AML patients, 46 AML patients in complete remission (CR) and 36 control samples. β-actin gene was used as internal reference. Relative changes of FANCG gene expression level were detected by 2(-ΔΔCT) method in newly diagnosed AML patients and control samples, in newly diagnosed AML and patient in CR, as well as in AML patients in CR and control samples. The results showed that the relative expression level of FANCG mRNA was 0.56 ± 0.27 in newly diagnosed group, 0.75 ± 0.54 in AML CR group, and 0.85 ± 0.45 in control group. The expression level of FANCG mRNA in newly diagnosed group was significantly lower than that in control and AML CR groups (P < 0.05). There was no statistically significant deference in comparison of AML CR group with the control group (P > 0.05). It is concluded that the expression of FANCG gene decrease in the newly diagnosed AML patients. There is no significant difference between AML CR group and control group, which indicated that FANCG gene may be related with the onset and the prognosis of AML, and may provide a clinical value for evaluating effect of chemotherapy.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Fanconi Anemia Complementation Group G Protein , Genetics , Leukemia, Myeloid, Acute , Genetics , Pathology , Prognosis , RNA, Messenger , Genetics , Real-Time Polymerase Chain ReactionABSTRACT
Human papillomavirus (HPV)-induced cervical cancer is the second most common cancer among women worldwide. Despite the encouraging development of the preventive vaccine for HPV, a vaccine for both prevention and therapy or pre-cancerous lesions remains in high priority. Thus far, most of the HPV therapeutic vaccines are focused on HPV E6 and E7 oncogene. However these vaccines could not completely eradicate the lesions. Recently, HPV E5, which is considered as an oncogene, is getting more and more attention. In this study, we predicted the epitopes of HPV16 E5 by bioinformatics as candidate peptide, then, evaluated the efficacy and chose an effective one to do the further test. To evaluate the effect of vaccine, rTC-1 (TC-1 cells infected by rAAV-HPV16E5) served as cell tumor model and rTC-1 loading mice as an ectopic tumor model. We prepared vaccine by muscle injection. The vaccine effects were determined by evaluating the function of tumor-specific T cells by cell proliferation assay and ELISPOT, calculating the tumor volume in mice and estimating the survival time of mice. Our in vitro and in vivo studies revealed that injection of E5 peptide+CpG resulted in strong cell-mediated immunity (CMI) and protected mice from tumor growth, meanwhile, prolonged the survival time after tumor cell loading. This study provides new insights into HPV16 E5 as a possible target on the therapeutic strategies about cervical cancer.
Subject(s)
Adult , Aged , Animals , Female , Humans , Mice , Middle Aged , Amino Acid Sequence , Cancer Vaccines , Allergy and Immunology , Cell Line , Cell Line, Tumor , Dependovirus , Genetics , Gene Expression Regulation, Viral , Allergy and Immunology , Genetic Vectors , Genetics , Human papillomavirus 16 , Genetics , Allergy and Immunology , Mice, Inbred C57BL , Neoplasms, Experimental , Allergy and Immunology , Virology , Oncogene Proteins, Viral , Genetics , Allergy and Immunology , Papillomavirus Infections , Allergy and Immunology , Virology , Papillomavirus Vaccines , Allergy and Immunology , Survival Analysis , T-Lymphocytes , Allergy and Immunology , Metabolism , Tumor Burden , Allergy and Immunology , Uterine Cervical Neoplasms , Allergy and Immunology , Virology , Vaccines, Subunit , Allergy and ImmunologyABSTRACT
Human papillomavirus (HPV)-induced cervical cancer is the second most common cancer among women worldwide. Despite the encouraging development of the preventive vaccine for HPV, a vaccine for both prevention and therapy or pre-cancerous lesions remains in high priority. Thus far, most of the HPV therapeutic vaccines are focused on HPV E6 and E7 oncogene. However these vaccines could not completely eradicate the lesions. Recently, HPV E5, which is considered as an oncogene, is getting more and more attention. In this study, we predicted the epitopes of HPV16 E5 by bioinformatics as candidate peptide, then, evaluated the efficacy and chose an effective one to do the further test. To evaluate the effect of vaccine, rTC-1 (TC-1 cells infected by rAAV-HPV16E5) served as cell tumor model and rTC-1 loading mice as an ectopic tumor model. We prepared vaccine by muscle injection. The vaccine effects were determined by evaluating the function of tumor-specific T cells by cell proliferation assay and ELISPOT, calculating the tumor volume in mice and estimating the survival time of mice. Our in vitro and in vivo studies revealed that injection of E5 peptide+CpG resulted in strong cell-mediated immunity (CMI) and protected mice from tumor growth, meanwhile, prolonged the survival time after tumor cell loading. This study provides new insights into HPV16 E5 as a possible target on the therapeutic strategies about cervical cancer.