作者：
Ling, H. Y.;Ou, H. S.;Lei, X. Y.;Zhu, B. Y.;Yin, W. D.;...
期刊：
RECENT ADVANCES IN CARDIOVASCULAR DISEASE: PROCEEDINGS OF THE 13TH WORLD CONGRESS HEART DISEASE,2007年:263-267
通讯作者：
Liao, D. F.
作者机构：
[Zhu, B. Y.; Yin, W. D.; Lei, X. Y.; Ou, H. S.; Liao, D. F.; Ling, H. Y.] Univ S China, Inst Pharm & Pharmacol, Div Pharmacoprote, Hengyang, Peoples R China.
通讯机构：
[Liao, D. F.] U;Univ S China, Inst Pharm & Pharmacol, Div Pharmacoprote, Hengyang, Peoples R China.
关键词：
MicroRNA;adipocyte;3T3-L1;differentiation;insulin signal
摘要：
This study investigated the MicroRNA expression profile during the differentiation from 3T3-L1 preadipocytes into adipocytes by miRNAs microarray analysis. We found both 3T3-L1 preadipocytes and adipocytes express 331 miRNAs. In adipocyte, 26 miRNAs were upregulated and 2 miRNAs were downregulated compared to 3T3-L1 preadipocytes. Interestingly, we observed that the target genes of miR-21, miR-298, let-7e were associated with insulin signal pathway.
通讯机构：
[Lei, Xiaoyong] U;Univ S China, Inst Pharm & Pharmacol, Hengyang 421001, Peoples R China.
关键词：
small interfering RNA;adenocarcinoma;Bcl-2;A549/DDP;apoptosis
摘要：
Bcl-2 is overexpressed in a variety of human tumors and is involved in tumorigenesis and chemoresistance. In this study, we investigated the inhibitory effect of the hairpin Bcl-2 small interfering (si)RNA on the expression of the Bcl-2 gene in the cisplatin (DDP)-resistant human lung adenocarcinoma cell line A549/DDP, and the effect of Bcl-2 siRNA on drug sensitization in A549/DDP cells. Bcl-2 siRNA and negative siRNA plasmids were constructed and stably transfected into A549/DDP cells. Reverse transcription-polymerase chain reaction, immunofluorescence microscopy and Western blot analysis were used to detect the target gene expression. Spontaneous cell apoptosis was detected by acridine orange and ethidium bromide staining. Drug sensitivity of the cells to DDP and diallyl disulfide (DADS) was analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Expression levels of Bcl-2 mRNA and protein in siRNA stable transfectants were clearly reduced compared with negative siRNA transfectants and untreated cells. MTT results indicated that Bcl-2 transfectants had a higher cell inhibition rate after treatment with 0.2-200 μg/ml DDP or 50-200 μM DADS. Flow cytometry revealed increased apoptosis in Bcl-2 siRNA cells. After the addition of 20 μg/ml DDP or 100 μM DADS, siRNA targeting of the Bcl-2 gene specifically down-regulated gene expression in A549/DDP cells, increased spontaneous apoptosis, and sensitized cells to DDP and DADS.
作者：
Ling, H. Y.;Ou, H. S.;Lei, X. Y.;Gao, Z. P.;Yin, W. D.;...
期刊：
RECENT ADVANCES IN CARDIOVASCULAR DISEASE: PROCEEDINGS OF THE 13TH WORLD CONGRESS HEART DISEASE,2007年:269-274
通讯作者：
Liao, D. F.
作者机构：
[Gao, Z. P.; Yin, W. D.; Lei, X. Y.; Ou, H. S.; Liao, D. F.; Ling, H. Y.] Univ S China, Inst Pharm & Pharmacol, Div Pharmacoprote, Hengyang, Peoples R China.
通讯机构：
[Liao, D. F.] U;Univ S China, Inst Pharm & Pharmacol, Div Pharmacoprote, Hengyang, Peoples R China.
关键词：
microRNA;adipocyte;3T3-L1;insulin resistance
摘要：
MicroRNAs (miRNAs) are a class of small non-coding RNAs which play an important role in metabolism of glucose and lipid in adipocyte. This study investigated miRNAs expression profiles during the formation process of insulin-resistant adipocyte derived from 3T3-L1 adipocyte. miRNAs were examined by miRNAs microarray analysis..The results showed that 71 miRNAs were upregulated and 33 miRNAs were downregulated during the formation of insulin resistance. Among them, miRNA (miR)-320, -494, -503, -202, -185 and -129-5p were significantly increased. Western blotting analysis showed that Anti-miR-320 increased GLUT4 expression.
摘要：
Bcl-XL is overexpressed in a variety of human tumors and is involved in tumorigenesis and chemoresistance. This study investigated the inhibitory effect of the hairpin Bcl-XL small interfering RNA (siRNA) on the expression of the Bcl-XL gene in the cisplatin (DDP)-resistant human lung adenocarcinoma cell line A549/DDP, and the effect of Bcl-XL siRNA on drug sensitization in A549/DDP cells. Bcl-XL siRNA and negative siRNA plasmids were constructed and stably transfected into A549/DDP cells. Reverse transcription-polymerase chain reaction and Western blot analysis were used to detect the target gene expression. Spontaneous apoptosis of cells was detected by acridine orange and ethidium bromide staining. Drug sensitivity of the cells to DDP was analyzed with dimethylthiazol-diphenyltetrazolium bromide (MTT) and flow cytometry. Expression levels of Bcl-XL mRNA and protein in siRNA stable transfectants were clearly reduced as compared with negative siRNA transfectants and untreated cells. MTT results indicated that Bcl-XL transfectants had a higher cell inhibition rate than the negative vector or untreated cells after treatment with 0.2–200 μg/ml DDP. Flow cytometry revealed increased apoptosis in Bcl-XL siRNA cells. After the addition of 20 μg/ml DDP, siRNA targeting of the Bcl-XL gene specifically down-regulated gene expression in A549/DDP cells, increased spontaneous apoptosis, and sensitized cells to DDP. The results showed that Bcl-XL siRNA contributed to an increase of DDP-induced cell death in non-small-cell lung cancer and sensitized cells to DDP, leading to increased the effectiveness of the drug in treating non-small-cell lung cancer.
摘要：
To investigate the changes in drug sensitivity of Bcl-2 siRNA transfected HepG2 cells. Bcl-2 siRNA and negative siRNA expression vector were constructed and stably transfected into HepG2 cells. RT-PCR and Immunofluorescence were used to detect the target gene expression. Western Blotting was used to detect Bcl-2, Bax and caspase-3 protein expressiom. Drug sensitivity of the cells to 5-fluorouracil (5-FU) and 10-hydroxycamptothecin (HCPT) were analyzed with MTT and flow cytometry. Results were following: (1) the mRNA and protein expression level of Bcl-2 in Bcl-2 siRNA stable transfectants were reduced compared with negative siRNA transfected or untreated cells. Accordingly, Bax protein expression had no change and caspase-3 protein expression showed significantly be up regulated; (2) MTT results showed that Bcl-2 siRNA transfectants had higher cell inhibitory rates after treated with 5-FU or HCPT; (3) flow cytometry results demonstrated that sub G1 population increased in Bcl-2 siRNA transfected cells compared with negative siRNA or untreated cells. After addition 5-FU (1300 mg/l) and HCPT (0.72 mg/l), Bcl-2 siRNA cells showed higher sub G1 population than negative siRNA or untreated cells. siRNA targeting Bcl-2 gene can specifically down-regulate Bcl-2 expression, increased Bax/Bcl-2 ratio expression and caspase-3 activity in HepG2 cells, which lead to increase cells spontaneous apoptosis and sensitize cells to 5-FU or HCPT. Bcl-2 siRNA may be a potential therapy agent against human hepatoblastoma.
作者机构：
[李强翔; 谢小英; 邓小金; 陈梅贵] Department of Endocrinology, Loudi Municipal Central Hospital, Loudi 417000 Hunan Province, China;[雷小勇] Department of Pharmacology, Nanhua University, Hengyang 421001 Hunan Province, China;[文格波; 欧玉兰] Institute of Clinical Medicine, Nanhua University, Hengyang 421001 Hunan Province, China
通讯机构：
[Li, Q.-X.] D;Department of Endocrinology, Loudi Municipal Central Hospital, China
作者机构：
[张洹] Institute of Hematolosy, Jinan University Medical College, Guangzhou 510632, China;Institute of Pharmacy and Pharmacology, Nanhua University, China;[雷小勇] Institute of Hematolosy, Jinan University Medical College, Guangzhou 510632, China, Institute of Pharmacy and Pharmacology, Nanhua University, China
通讯机构：
[Huan Zhang] I;Institute of Hematology, Jinan University Medical College, Guangzhou
摘要：
Objective: To investigate the effect of two antisense oligonucleotides on cell surviving, bcl-2 expression and apoptosis of leukemia cells. Methods: The experimental assays were performed with cell culture, immunochemistry and flowcytometry. Results: The two antisense oligodeoxynucleotides, combined with Vp16 or Ara-c or DNR, were able to decline the survival rate of myeleukemic cells, downregulate bcl-2 gene expression and induce apoptosis of leukemic cells significantly, as compared with Vp16 or Ara-c or DNR alone. Conclusion: It is possible for the two new bcl-2 antisenses to be developed into clinical trials for leukemia and tumor with bcl-2 gene overexpression.
摘要：
To investigate the inhibitory effect of the Bcl-XL small interfering RNA (siRNA) on Bcl-XL gene expression in the human gastric cancer cell line MGC-803, green fluorescent protein (GFP) siRNA was constructed and transfected into MGC-803 cells, together with GFP expression vector pTrace SV40. GFP expression levels were observed using fluorescence microscopy. Bcl-XL siRNA and negative siRNA were then constructed and stably transfected into MGC-803 cells. RT-PCR and immunofluorescence were used to detect the expression of Bcl-XL. Spontaneous apoptosis was detected by acridine orange (AO) and flow cytometry. Results were as follows: (1) 48 h after GFP expression vector and GFP siRNA co-transfection, the expression level of GFP in the GFP siRNA group was much lower than the negative siRNA group, according to fluorescence microscopy results. The mRNA and protein levels of Bcl-XL in Bcl-XL siRNA stable transfectants were reduced to almost background level compared with negative siRNA transfectants or untreated cells. (2) Changes in nucleus morphology was observed by AO staining nucleic and flow cytometry analysis, which showed that stable Bcl-XL siRNA transfectants have an increased spontaneous apoptosis (21.17%±1.26% vs. 1.19%±0.18% and 1.56%±0.15% respectively, P<0.05 vs. negative siRNA or untreated control). siRNA targeting GFP or Bcl-XL genes can specifically suppress GFP or Bcl-XL expression in MGC-803 cells, and Bcl-XL siRNA can increase spontaneous apoptosis. Bcl-XL siRNA may be a beneficial agent against human gastric adenocarcinoma.
