1.The expression of miR-199a-3p in breast cancer and its effect
Dachang MA ; Cheng CHEN ; Duoming WU ; Honglei WANG ; Li WU
China Oncology 2016;26(6):481-486
Background and purpose:Multiple microRNAs (miRNAs) are abnormally expressed in breast cancer and play an important role in the regulation of breast cancer. miRNAs may be a new target for the treatment of breast cancer. This study aimed to investigate the expression of miR-199a-3p in breast cancer and the effect of miR-199a-3p on proliferation and apoptosis of breast cancer.Methods:Real-time PCR was used to test the expression of miR-199a-3p in breast cancer tissues, normal breast tissues, breast cancer cells and normal breast cells. Overexpression (or silencing the expression) of miR-199a-3p was conducted by transfecting MDA-MB-231 with miR-199a-3p mimics (or inhibitors). The proliferation of MDA-MB-231 was detected by MTT method. The apoptosis of MDA-MB-231 was investigated by Hoechst staining and caspase-3 activity assay kit.Results:Compared to corresponding non-tumor breast tissues (or normal breast cell HBL-100), lower levels of miR-199a-3p were expressed in breast cancer tissues or breast cancer cells. Overexpression of miR-199a-3p induced by miR-199a-3p mimic inhibited the proliferation and promoted the apoptosis of MDA-MB-231, while silencing the expression of miR-199a-3p induced by miR-199a-3p in-hibitor increased the proliferation and suppressed the apoptosis of MDA-MB-231.Conclusion:The expression of miR-199a-3p is lower in breast cancer, which shows its tumor suppression effect by regulating the proliferation and apoptosis of breast cancer cells.
2.The A-204C Polymorphism in CYP7A1 Gene Affects Its Promoter Activity
Yujuan CHEN ; Sizhong ZHANG ; Cuiying XIAO ; Dachang TAO ; Guoping HE ; Yingcheng WANG ; Yunqiang LIU ; Yongxin MA
Chinese Journal of Biochemistry and Molecular Biology 2006;22(6):450-453
cholesterol 7α-hydroxylase gene ( CYP7A 1 ) plays a key role in the catabolism of cholesterol into bile acids. To investigate whether the A-204C polymorphism in CYP7A1 gene affects the gene expression,using luciferase as the reporter gene, four recombinants were constructed by inserting forward or reverse sequence with A or C allele at the polymorphism site into the promoter-less vector pGL3-basic. The constructs were then transfected into four cell lines and the luciferase activity of each expression vector was examined by dual luciferase reporter gene assay system. The results showed that activities of the forward sequence of both genotypes were higher than that of reverse sequence. Promoter activity of the recombinants with A allele was about one third lower than that with C allele. According to the analysis with TRANSFAC database, there may exist a Zic3 binding site when there is the C allele at -204. Our study indicates that the A-204 C polymorphism in CYP7A1 promoter region decreases its promoter activity and thus represses the gene expression, possibly due to the lack of a potential Zic3 binding site.
3.Identification of proteins interacting with the circadian clock protein PER1 in tumors using bacterial two-hybrid system technique.
Yu ZHANG ; Youlin YAO ; Siyuan JIANG ; Yilu LU ; Yunqiang LIU ; Dachang TAO ; Sizhong ZHANG ; Yongxin MA
Chinese Journal of Medical Genetics 2015;32(2):192-197
OBJECTIVETo identify protein-protein interaction partners of PER1 (period circadian protein homolog 1), key component of the molecular oscillation system of the circadian rhythm in tumors using bacterial two-hybrid system technique.
METHODSHuman cervical carcinoma cell Hela library was adopted. Recombinant bait plasmid pBT-PER1 and pTRG cDNA plasmid library were cotransformed into the two-hybrid system reporter strain cultured in a special selective medium. Target clones were screened. After isolating the positive clones, the target clones were sequenced and analyzed.
RESULTSFourteen protein coding genes were identified, 4 of which were found to contain whole coding regions of genes, which included optic atrophy 3 protein (OPA3) associated with mitochondrial dynamics and homo sapiens cutA divalent cation tolerance homolog of E. coli (CUTA) associated with copper metabolism. There were also cellular events related proteins and proteins which are involved in biochemical reaction and signal transduction-related proteins.
CONCLUSIONIdentification of potential interacting proteins with PER1 in tumors may provide us new insights into the functions of the circadian clock protein PER1 during tumorigenesis.
Base Sequence ; Cell Line, Tumor ; Escherichia coli ; genetics ; metabolism ; Humans ; Molecular Sequence Data ; Neoplasms ; genetics ; metabolism ; Period Circadian Proteins ; genetics ; metabolism ; Protein Binding ; Proteins ; genetics ; metabolism ; Two-Hybrid System Techniques
4.Investigation and Analysis of Medication Information Labeling in 157 Package Inserts of Anticancer Drugs
Jing MA ; Yan WANG ; Bo LU ; Shouxiang TANG ; Lihua YUAN ; Xuemin ZHOU
China Pharmacy 2019;30(7):985-990
OBJECTIVE: To investigate and analyze medication information labeling in package inserts of anticancer drugs, and to provide reference for clinical rational use. METHODS: The package inserts of anticancer drugs were collected from drug catalogues of 3 Third Grade Class A hospitals in Nanjin. Common problems of drug package inserts (whether the main contents arweree contradictory or not and whether the contents were fully expressed, etc.), complete specific labeling items (detailed contents of “ADR” “contraindication” “precautions” and other items), detailed intravenous injection dispensing guidance (solvent selection, precautions during dispensing, etc.), package insert labeling difference of drugs with same general name and route of administration were evaluated according to Drug Package Inserts and Label Management Regulation,Regulations for Chemical Drugs and Biological Products for Treatment. RESULTS: A total of 157 package inserts for anticancer drugs were collected and divided into domestic drugs (80 pieces) and imported drugs (77 pieces) according to the source as well as also divided into oral preparation (44 pieces) and injection (113 pieces). The common problems of package inserts for anticancer drugs contained contradictory main contents, incomplete description, Chinese character errors, missing items and simple description of drug interactions, etc. Compared with domestic or oral anticancer drugs, the labeling rate of each item in the import or injection anticancer drug package inserts was higher, but specific labeling items such as prevention and treatment of vomiting (<20%) under “precautions” and interference of drugs on clinical tests (<40%) were lower. The labeling rate of serious ADR after large dose or long-term use was all less than 41% under the item of “drug overdose” (except for imported drugs). The labeling rate of intravenous dispensing guidance of imported anticancer drug injection package inserts about preparations was higher than that of domestic ones. There were differences in labeling items as “precautions” (30/56,53.57%), “pharmacological toxicology” (29/56,51.79%), “contraindication” (26/56,46.43%) among 56 groups of drug package inserts with same general name and route of administration. CONCLUSIONS: The labeling items for drug package inserts of anticancer drugs need to be further standardized and improved. It is recommended that the relevant departments force pharmaceutical manufacturers to regularly supplement the deficiencies in the package inserts to improve the safety of drug use in clinic.