This is first report about the simultaneous over-expression of both Insulin-like growth factor (IGF- I ) and its receptor (IGF- I R) at mRNA level in human primary hepatic Cancer (PHC). In 10 PHC samples from China, I...This is first report about the simultaneous over-expression of both Insulin-like growth factor (IGF- I ) and its receptor (IGF- I R) at mRNA level in human primary hepatic Cancer (PHC). In 10 PHC samples from China, IGF-I and IGF- I R were both over-expressed, whereas only a background signal was detected in normal liver. In 5 pairs of PHC and its non- tumorous adjacent liver tissues from South Africa, IGF- I and IGF- I R were also over-expressed in PHC. mRNA expression of IGF- I in all 5 cases and IGF- I R in 4 of 5 cases were higher in cancer than non- tumorous adjacent liver tissues. These results strongly implicate that an autocrine and/ or paracrine mechanism might be Involved in formation and progression of PHC.展开更多
Variations in expression of insulin-like growth factors （IGFs） system genes in rat ovarian and uterine tissues were investigated by real-time quantitative PCR amplification using RT2 ProfilerTM PCR Array Rat Insulin...Variations in expression of insulin-like growth factors （IGFs） system genes in rat ovarian and uterine tissues were investigated by real-time quantitative PCR amplification using RT2 ProfilerTM PCR Array Rat Insulin Signaling Pathway （PARN-030A） chips. According to results of gene detection, a total of 15 differeutially expressed genes were screened in ovarian tissues, including 3 upregulated genes and 10 downregulated genes; a total of 22 differentially expressed genes were screened in uterine tissues, including 5 upregulated genes and 17 downregulated genes. These differentially expressed genes might be related to the insulin resistance.展开更多
Objective： To investigate the effect of insulin-like growth factor 1 receptor （IGF1R） small interfering RNA （siRNA） on the growth of human liver cancer SMMC7721 cell xenograft in nude mice. Methods： siRNA target...Objective： To investigate the effect of insulin-like growth factor 1 receptor （IGF1R） small interfering RNA （siRNA） on the growth of human liver cancer SMMC7721 cell xenograft in nude mice. Methods： siRNA targeting IGF1R was designed, and plasmid SMMC7721-1GF1R-siRNA was constructed and transfected into SMMC7721 cells （SMMC7721-1GF1R-siRNA cells）; the cells transfected with SMMC7721-1GF1 R-mutation （SMMC7721-1GF1 R-mutation cells） were used as negative con- trol, and untransfected cells as empty control. Stable cell clones were screened by G418, and transplanted into nude mice to establish cancer xenograft. Tumor growth was monitored. Tumor morphology was observed with HE staining. The expression of IGF1R protein in tumor tissues was detected by Western blot. Microvessel density （MVD） in tumor tissues was detected by SP immunohistochemistry. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL） assay. Results： The tumor volume was significantly smaller in SMMC7721-1GF1R-siRNA group than in SMMC7721-1GF1R-mutation and SMMC7721 groups （P 〈 0.05）. Necrosis and cell apoptosis were found in SMMC7721- IGF1R-siRNA group. The expression of IGF1R protein was significantly lower in SMMC7721-1GF1R-siRNA group than in SMMC7721-1GF1R-mutation and SMMC7721 groups （P 〈 0.05）. MVD was significantly lower in SMMC7721-1GF1R-siRNA group than in SMMC7721-1GF1R-mutation and SMMC7721 groups （11.3 ± 4.4 vs. 36.7 ± 7.6 and 28.4 ±6.5, P 〈 0.05）. The apoptosis rate of tumor cells was significantly higher in SMMC7721-1GF1R-siRNA group than in SMMC7721-1GF1R-mutation and SMMC7721 groups [（50.2 ± 6.4）% vs. （5.4 ± 1.0）% or （6.0 ±2.1）%, P〈0.05]. Conclusion： IGF1R siRNA can inhibit the growth of SMMC7721 cell xenograft in nude mice.展开更多
Following extensive bowel resection, the intestinal tract undergoes a variety of adaptive responses to enhance bowel function. The purpose of this study was to determine the effect of glutamine-supplemented parenteral...Following extensive bowel resection, the intestinal tract undergoes a variety of adaptive responses to enhance bowel function. The purpose of this study was to determine the effect of glutamine-supplemented parenteral nutrition on mucosal cellularity and gut function. In addition, enterocyte gene expression of two relevant systems was also characterized and related to the structural and functional changes that occurred. Male Wistar rats underwent a 60% small bowel resection and jugular vein catheterization and were randomized into two groups. The control group (n = 10) received a standard intravenous nutritional solution and the study group (n = 10) received a similar solution but enriched with alanylglutamine dipeptide. After 7 days blood was taken for amino acid analysis, and bowel was harvested to determine mucosal morphology and expression of mucosal cell glutaminase and IGF-I mRNA. Mesentery lymphnodes were cultured to determine the presence of bacteria and thus access bacteria translocation. Serum glutamine concentration and mucosal architecture were maintained in the study group compared to the controls. Seventy percent of lymphnodes were cultured positive in control vs. only 20% in the study group (P展开更多
INIRODUCTIONAccording to the therapeutic effect and strategy ofantisense RNA for hepatoccllular carcinoma(HCC),we have specifically synthesized partialcDNA of human insulin-like growth factor Ⅱ(IGF-Ⅱ)and constructed...INIRODUCTIONAccording to the therapeutic effect and strategy ofantisense RNA for hepatoccllular carcinoma(HCC),we have specifically synthesized partialcDNA of human insulin-like growth factor Ⅱ(IGF-Ⅱ)and constructed IGF-Ⅱ cDNA antisenseeukaryotic expression vector.The constructedvector was introduced into hepatoma cell lineSMMC-7721 to block the intrinsic IGF-Ⅱexpression.The biological behavior changes ofhepatoma cells were observed.All these展开更多
A major problem which is poorly understood in the management of bladder cancer is low sensitivity to chemotherapy and high recurrence after transurethral resection. Insulin-like growth factor 1 receptor (IGF-1R) signa...A major problem which is poorly understood in the management of bladder cancer is low sensitivity to chemotherapy and high recurrence after transurethral resection. Insulin-like growth factor 1 receptor (IGF-1R) signaling plays a very important role in progression, invasion and metastasis of bladder cancer cells. In this study, we investigated whether IGF-1R was involved in the growth stimulating activity and drug resistance of bladder cancer cells. The results showed: The mRNAs of IGF-1, IGF-2 and IGF-1R were strongly expressed in serum-free cultured T24 cell line, whereas normal urothelial cells did not express these factors/receptors or only in trace levels; T24 cell responded far better to growth stimulation by IGF-1 than did normal urothelial cells; blockage of IGF1R by antisense oligodeoxynucleotide (ODN) significantly inhibited the growth of T24 cell and enhanced sensitivity and apoptosis of T24 cells to mitomycin (MMC). These results suggested that blockage of IGF-IR signaling might potentially contribute to the treatment of bladder cancer cells which are insensitive to chemotherapy.展开更多
The DNA sequence encoding the chicken＇s insulin-like growth factor Ⅰ （IGF-Ⅰ） was amplified with the reverse transcription polymerase chain reaction （RT-PCR）, which was then cloned into vector pMD18-T and sequen...The DNA sequence encoding the chicken＇s insulin-like growth factor Ⅰ （IGF-Ⅰ） was amplified with the reverse transcription polymerase chain reaction （RT-PCR）, which was then cloned into vector pMD18-T and sequenced. The sequencing result showed that there was 100% homology among the documented sequences and the sequence reported in this article, which was successfully inserted into the expressing plasmid pRLC and was highly expressed in E.coli. The Tricine-SDS- PAGE result showed that the cloned recombinant protein was expressed in the form of inclusion bodies in the E.coli cell with molecular weight of 7.6 kD and was amount to 23% of the whole protein in the E.coli cell. Western blotting indicated that recombinant protein had the antigenicity of IGF- Ⅰ. The inclusion bodies were subsequently dissolved in 7 M guanidine chloride and renatured with dilution in refolding buffer containing 0.5 M arginine. To obtain pure protein, the renatured chicken IGF- Ⅰ was desalting by Hiprep 26/10 and purified by Hiprep Sephacryl S-200 chromatography. The biological activities of IGF- Ⅰ product were assayed in NIH 3T3 cells and osteoblastic cells of embryonic chicken by using MTT method. The results show that the expressed IGF- Ⅰ can obviously stimulate NIH3T3 cells and osteoblastic cells to proliferate at the concentration ranging from 100, 200, 400 to 800 ng mL^-1, suggesting that the protein has its biological activities.展开更多
本试验采用单核苷酸多态性(single nucleotide polymorphism,SNP)直接测序法检测猪锚蛋白重复序列和SOCS盒蛋白基因4(ankyrin repeat and SOCS box containing protein 4,Asb4)与生长因子受体结合蛋白基因10(growth factor recep-tor-bo...本试验采用单核苷酸多态性(single nucleotide polymorphism,SNP)直接测序法检测猪锚蛋白重复序列和SOCS盒蛋白基因4(ankyrin repeat and SOCS box containing protein 4,Asb4)与生长因子受体结合蛋白基因10(growth factor recep-tor-bound protein 10,Grb10)在不同组织器官中的印迹状态。首先,克隆得到了1350bp的Asb4基因cDNA序列及1811bp的Grb10基因cDNA序列,然后进行了SNP直接测序法检测。结果发现,Asb4在1月龄仔猪所有检测组织器官中均为双等位基因表达,而Grb10在1月龄仔猪的舌、肾脏、胃、小肠和脑中为父源等位基因表达,在其他组织器官中为双等位基因表达。实时定量PCR结果表明,Grb10在10种组织器官中的表达量存在显著性差异(P<0.05),其中在肺脏组织中的表达量最高,在小肠和脑中表达量最低。上述结果表明,Grb10可能是猪父源表达的印迹基因,而Asb4则属于猪非印迹基因。展开更多
文摘This is first report about the simultaneous over-expression of both Insulin-like growth factor (IGF- I ) and its receptor (IGF- I R) at mRNA level in human primary hepatic Cancer (PHC). In 10 PHC samples from China, IGF-I and IGF- I R were both over-expressed, whereas only a background signal was detected in normal liver. In 5 pairs of PHC and its non- tumorous adjacent liver tissues from South Africa, IGF- I and IGF- I R were also over-expressed in PHC. mRNA expression of IGF- I in all 5 cases and IGF- I R in 4 of 5 cases were higher in cancer than non- tumorous adjacent liver tissues. These results strongly implicate that an autocrine and/ or paracrine mechanism might be Involved in formation and progression of PHC.
基金Supported by National Natural Science Foundation of China(30973927)
文摘Variations in expression of insulin-like growth factors （IGFs） system genes in rat ovarian and uterine tissues were investigated by real-time quantitative PCR amplification using RT2 ProfilerTM PCR Array Rat Insulin Signaling Pathway （PARN-030A） chips. According to results of gene detection, a total of 15 differeutially expressed genes were screened in ovarian tissues, including 3 upregulated genes and 10 downregulated genes; a total of 22 differentially expressed genes were screened in uterine tissues, including 5 upregulated genes and 17 downregulated genes. These differentially expressed genes might be related to the insulin resistance.
文摘Objective： To investigate the effect of insulin-like growth factor 1 receptor （IGF1R） small interfering RNA （siRNA） on the growth of human liver cancer SMMC7721 cell xenograft in nude mice. Methods： siRNA targeting IGF1R was designed, and plasmid SMMC7721-1GF1R-siRNA was constructed and transfected into SMMC7721 cells （SMMC7721-1GF1R-siRNA cells）; the cells transfected with SMMC7721-1GF1 R-mutation （SMMC7721-1GF1 R-mutation cells） were used as negative con- trol, and untransfected cells as empty control. Stable cell clones were screened by G418, and transplanted into nude mice to establish cancer xenograft. Tumor growth was monitored. Tumor morphology was observed with HE staining. The expression of IGF1R protein in tumor tissues was detected by Western blot. Microvessel density （MVD） in tumor tissues was detected by SP immunohistochemistry. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL） assay. Results： The tumor volume was significantly smaller in SMMC7721-1GF1R-siRNA group than in SMMC7721-1GF1R-mutation and SMMC7721 groups （P 〈 0.05）. Necrosis and cell apoptosis were found in SMMC7721- IGF1R-siRNA group. The expression of IGF1R protein was significantly lower in SMMC7721-1GF1R-siRNA group than in SMMC7721-1GF1R-mutation and SMMC7721 groups （P 〈 0.05）. MVD was significantly lower in SMMC7721-1GF1R-siRNA group than in SMMC7721-1GF1R-mutation and SMMC7721 groups （11.3 ± 4.4 vs. 36.7 ± 7.6 and 28.4 ±6.5, P 〈 0.05）. The apoptosis rate of tumor cells was significantly higher in SMMC7721-1GF1R-siRNA group than in SMMC7721-1GF1R-mutation and SMMC7721 groups [（50.2 ± 6.4）% vs. （5.4 ± 1.0）% or （6.0 ±2.1）%, P〈0.05]. Conclusion： IGF1R siRNA can inhibit the growth of SMMC7721 cell xenograft in nude mice.
文摘Following extensive bowel resection, the intestinal tract undergoes a variety of adaptive responses to enhance bowel function. The purpose of this study was to determine the effect of glutamine-supplemented parenteral nutrition on mucosal cellularity and gut function. In addition, enterocyte gene expression of two relevant systems was also characterized and related to the structural and functional changes that occurred. Male Wistar rats underwent a 60% small bowel resection and jugular vein catheterization and were randomized into two groups. The control group (n = 10) received a standard intravenous nutritional solution and the study group (n = 10) received a similar solution but enriched with alanylglutamine dipeptide. After 7 days blood was taken for amino acid analysis, and bowel was harvested to determine mucosal morphology and expression of mucosal cell glutaminase and IGF-I mRNA. Mesentery lymphnodes were cultured to determine the presence of bacteria and thus access bacteria translocation. Serum glutamine concentration and mucosal architecture were maintained in the study group compared to the controls. Seventy percent of lymphnodes were cultured positive in control vs. only 20% in the study group (P
基金the National Natural Science Foundation of Guangdong Province,No.940319.
文摘INIRODUCTIONAccording to the therapeutic effect and strategy ofantisense RNA for hepatoccllular carcinoma(HCC),we have specifically synthesized partialcDNA of human insulin-like growth factor Ⅱ(IGF-Ⅱ)and constructed IGF-Ⅱ cDNA antisenseeukaryotic expression vector.The constructedvector was introduced into hepatoma cell lineSMMC-7721 to block the intrinsic IGF-Ⅱexpression.The biological behavior changes ofhepatoma cells were observed.All these
文摘A major problem which is poorly understood in the management of bladder cancer is low sensitivity to chemotherapy and high recurrence after transurethral resection. Insulin-like growth factor 1 receptor (IGF-1R) signaling plays a very important role in progression, invasion and metastasis of bladder cancer cells. In this study, we investigated whether IGF-1R was involved in the growth stimulating activity and drug resistance of bladder cancer cells. The results showed: The mRNAs of IGF-1, IGF-2 and IGF-1R were strongly expressed in serum-free cultured T24 cell line, whereas normal urothelial cells did not express these factors/receptors or only in trace levels; T24 cell responded far better to growth stimulation by IGF-1 than did normal urothelial cells; blockage of IGF1R by antisense oligodeoxynucleotide (ODN) significantly inhibited the growth of T24 cell and enhanced sensitivity and apoptosis of T24 cells to mitomycin (MMC). These results suggested that blockage of IGF-IR signaling might potentially contribute to the treatment of bladder cancer cells which are insensitive to chemotherapy.
基金This work was supported by the National Natural Science Foundation of China(30270998) Natural Science Foundation of Jiangsu Province,China(Q200443).
文摘The DNA sequence encoding the chicken＇s insulin-like growth factor Ⅰ （IGF-Ⅰ） was amplified with the reverse transcription polymerase chain reaction （RT-PCR）, which was then cloned into vector pMD18-T and sequenced. The sequencing result showed that there was 100% homology among the documented sequences and the sequence reported in this article, which was successfully inserted into the expressing plasmid pRLC and was highly expressed in E.coli. The Tricine-SDS- PAGE result showed that the cloned recombinant protein was expressed in the form of inclusion bodies in the E.coli cell with molecular weight of 7.6 kD and was amount to 23% of the whole protein in the E.coli cell. Western blotting indicated that recombinant protein had the antigenicity of IGF- Ⅰ. The inclusion bodies were subsequently dissolved in 7 M guanidine chloride and renatured with dilution in refolding buffer containing 0.5 M arginine. To obtain pure protein, the renatured chicken IGF- Ⅰ was desalting by Hiprep 26/10 and purified by Hiprep Sephacryl S-200 chromatography. The biological activities of IGF- Ⅰ product were assayed in NIH 3T3 cells and osteoblastic cells of embryonic chicken by using MTT method. The results show that the expressed IGF- Ⅰ can obviously stimulate NIH3T3 cells and osteoblastic cells to proliferate at the concentration ranging from 100, 200, 400 to 800 ng mL^-1, suggesting that the protein has its biological activities.
文摘本试验采用单核苷酸多态性(single nucleotide polymorphism,SNP)直接测序法检测猪锚蛋白重复序列和SOCS盒蛋白基因4(ankyrin repeat and SOCS box containing protein 4,Asb4)与生长因子受体结合蛋白基因10(growth factor recep-tor-bound protein 10,Grb10)在不同组织器官中的印迹状态。首先,克隆得到了1350bp的Asb4基因cDNA序列及1811bp的Grb10基因cDNA序列,然后进行了SNP直接测序法检测。结果发现,Asb4在1月龄仔猪所有检测组织器官中均为双等位基因表达,而Grb10在1月龄仔猪的舌、肾脏、胃、小肠和脑中为父源等位基因表达,在其他组织器官中为双等位基因表达。实时定量PCR结果表明,Grb10在10种组织器官中的表达量存在显著性差异(P<0.05),其中在肺脏组织中的表达量最高,在小肠和脑中表达量最低。上述结果表明,Grb10可能是猪父源表达的印迹基因,而Asb4则属于猪非印迹基因。
