Reactivation of the insulin-like growth factor-Ⅱsignaling pathway in human hepatocellular carcinoma

Constitutive activation of the insulin-like growth factor (IGF)-signaling axis is frequently observed in human hepatocellular carcinoma(HCC).Especially the over- expression of the fetal growth factor IGF-Ⅱ,IGF-Ⅰ receptor(IGF-IR),and cytoplasmic downstream effectors such as insulin-receptor substrates(IRS)contribute to proliferation,anti-apoptosis,and invasive behavior. This review focuses on the relevant alterations in this signaling pathway and independent in vivo models that support the central role IGF-Ⅱsignaling during HCC development and progression.Since this pathway has become the center of interest as a target for potential anti-cancer therapy in many types of malignancies,various experimental strategies have been developed,including neutralizing antibodies and selective receptor kinase inhibitors,with respect to the specific and efficient reduction of oncogenic IGF-Ⅱ/IGF-IR-signaling.

Abnormal expression of insulin-like growth factor-Ⅱ and its dynamic quantitative analysis at different stages of hepatocellular carcinoma development

BACKGROUND:Hepatocellular carcinoma(HCC)is characterized by multiple causes,clear multiple stages and a multifocal process of tumor progression related intimately to the overexpression of many cellular factors. The aim of this study was to investigate the dynamic expression of insulin-like growth factor-Ⅱ(IGF-Ⅱ) and its abnormal alteration in the early stages of HCC development. METHODS:Hepatoma models were induced by 2-fluorenylacetamide(2-FAA)in male Sprague-Dawley rats.Morphological changes of rat livers were assessed by pathological examination(HE staining).The levels of IGF-Ⅱexpression in the livers and sera of rats were quantitatively detected by an enzyme-linked immunosorbent assay(ELISA).Simultaneously,the expression and cellular distribution of liver IGF-Ⅱwere analyzed by immunohistochemistry. RESULTS:Histological examination confirmed that rat hepatocytes showed changes from granule-like degeneration to atypical hyperplasia to HCC,and progressively increasing hepatic IGF-Ⅱlevels during HCC development.The levels of hepatic or serum IGF- Ⅱin HCC tissues and sera were significantly higher than those in normals and rats with degeneration.The immunohistochemical evidence indicated the positiveexpression and hepatocyte distribution of IGF-Ⅱin rat hepatoma.A positive relationship of IGF-Ⅱlevels was found between liver tissues and sera of experimental rats (P【0.01). CONCLUSION:Hepatic IGF-Ⅱmay participate in hepatocyte cancer development and detection of IGF-Ⅱ expression during HCC development could be a useful molecular marker for its early diagnosis.

Interplay between micro RNA-17-5p, insulin-like growth factor-Ⅱ through binding protein-3 in hepatocellular carcinoma

AIM: To investigate the effect of microR NA on insulinlike growth factor binding protein-3(IGFBP-3) and hence on insulin-like growth factor-Ⅱ(IGF-Ⅱ) bioavailability in hepatocellular carcinoma(HCC).METHODS: Bioinformatic analysis was performed using microrna.org, DIANA lab and Segal lab softwares. Total RNA was extracted from 23 HCC and 10 healthy liver tissues using mir Vana mi RNA Isolation Kit. microR NA-17-5p(miR-17-5p) expression was mimicked and antagonized in Hu H-7 cell lines using Hi Per Fect Transfection Reagent, then total RNA was extracted using Biozol reagent then reverse transcribed into cD NA followed by quantification of mi R-17-5p and IGFBP-3 expression using Taq Man real-time quantitative PCR. Luciferase reporter assay was performed to validate the binding of miR-17-5p to the 3'UTR of IGFBP-3. Free IGF-Ⅱ protein was measured in transfected Hu H-7 cells using IGF-Ⅱ ELISA kit. RESULTS: Bioinformatic analysis revealed IGFBP-3 as a potential target for miR-17-5p. Screening of miR-17-5p and IGFBP-3 revealed a moderate negative correlation in HCC patients, where mi R-17-5p was extensively underexpressed in HCC tissues(P = 0.0012), while IGFBP-3 showed significant upregulation in the same set of patients(P = 0.0041) compared to healthy donors. Forcing mi R-17-5p expression in Hu H-7 cell lines showed a significant downregulation of IGFBP-3 mR NA expression(P = 0.0267) and a significant increase in free IGF-Ⅱ protein(P = 0.0339) compared to mock untransfected cells using unpaired t-test. Luciferase assay validated IGFBP-3 as a direct target of mi R-17-5p; luciferase activity was inhibited by 27.5% in cells co-transfected with miR-17-5p mimics and the construct harboring the wild-type binding region 2 of IGFBP-3 compared to cells transfected with this construct alone(P = 0.0474).CONCLUSION: These data suggest that regulating IGF-Ⅱ bioavailability and hence HCC progression can be achieved through targeting IGFBP-3 via manipulating the expression of miR NAs.

Changes of insulin-like growth factor-Ⅱ and insulin-like growth factor binding protein-3 in cerebrospinal fluid of children with tuberculous meningitis

BACKGROUND： Recent studies have found that insulin-like growth factors （IGFs） and insulin-like growth factor binding protein-3 （IGFBP-3） have stronger neurotrophic and neuroprotective effects. But whether their levels in cerebrospinal fluid could be used as an auxiliary indicator in differentially diagnosing tuberculous meningitis and viral encephalitis is not yet clear. OBJECTIVE： To explore the changes of insulin-like growth factor-Ⅱ （IGF-Ⅱ ） and IGFBP-3 in cerebrospinal fluid （CSF） of children with tuberculous meningitis and the significance of the changes. DESIGN： A non-randomized concurrent controlled study. SETTING： Department of Pediatric Internal Medicine, the First Affiliated Hospital of Xinxiang Medical College. PARTICIPANTS： Thirty children with tuberculous meningitis （14 males and 16 females） were selected from the Department of Pediatric Internal Medicine, the First Affiliated Hospital of Xinxiang Medical College from January 2005 to December 2006. Tuberculous meningitis was diagnosed according to their clinical manifestations, the history of close contact with tuberculosis, typical cerebrospinal fluid changes of tuberculous meningitis, positive tuberculosis antibody and effective antituberculosis treatment. There were 30 children （13 males and 17 females） with viral encephalitis, and viral encephalitis was diagnosed according to epidemiological history, clinical manifestations, conventional and biochemical changes of cerebrospinal fluid, and negative bacteriology judgment. Meanwhile, 30 children （13 males and 17 females） without infectious and central nervous system disease were selected as the control group. Informed consent was obtained from the parents of all the enrolled children. METHODS： ①The lumbar puncture operation was implemented immediately to obtain cerebrospinal fluid （3 mL）. The contents of IGF-Ⅱ and IGFBP-3 were detected with immunoradiometric assay. The concentrations of glucose and protein in cerebrospinal fluid were determined with a dry-chemical method. The number of white blood cells was counted by Fushi Method. ②The Pearson correlation analysis was used to analyze the correlation of the contents of IGF-Ⅱ and IGFBP-3 in cerebrospinal fluid with the leucocyte counting and the concentrations of glucose and protein in cerebrospinal fluid. MAIN OUTCOME MEASURES： The contents of IGF- Ⅱ and IGFBP-3 in cerebrospinal fluid, and their correlation with the leucocyte counting and the concentrations of glucose and protein in cerebrospinal fluid. RESULTS： ①Contents of IGF-Ⅱ and IGFBP-3 in cerebrospinal fluid： The contents of IGF-Ⅱ and IGFBP-3 in cerebrospinal fluid in the tuberculous meningitis group were significantly higher than those in the encephalitis virus group and control group （P 〈 0.05）. There was no significant difference in the contents of IGF- Ⅱ and IGFBP-3 in cerebrospinal fluid between the viral encephalitis group and control group （P 〉 0.05）. ②Correlation： The IGF- Ⅱ and IGFBP-3 contents in cerebrospinal fluid were positively correlated with the protein concentration in cerebrospinal fluid （r =0.821, 0.855, P 〈 0.01）, but negatively with the glucose （r =0.742, - 0.605, P 〈 0.01）. CONCLUSION- ①IGFs and IGVBPs are involved in the pathophysiological process of tuberculous meningitis, as well as the glucose and protein metabolism in cerebrospinal fluid. ②The IGF-Ⅱ and IGFBP-3 contents in cerebrospinal fluid can be used as the auxiliary indicators to differentially diagnose tuberculous meningitis and viral enceohalitis.

Expression of insulin-like growth factor Ⅱ and its receptor in hepatocellular carcinogenesis

INTRODUCTIONInsulin-like growth factor Ⅱ(IGF-Ⅱ) is a mitogenic peptide of 74 kD and is mostly synthesized in fetal liver tissue .IGF-Ⅱ is believed to play an important role in fetal growth and development and is involved in cellular proliferation and differentiation[1-5]. Recently ,several researchers have reported increased expression of the IGF-Ⅱgene in human hepatocellular carcinoma (HCC) and adjacent non-cancerous liver tissues [6-10].

The immunocytochemical localization of insulin-like growth factor Ⅱ in human cirrhosis

Sections of 30 cases of human cirrhosis were stained with rabbit anti-insulin-likegrowth factor Ⅱ(IGF Ⅱ)by double PAP method.By the serological examination 15 patientsshowed HBV infection and sections of 14 eases were HBsAg postively with a total rate of 67％(20 cases).The IGF Ⅱ was positive in the cytoplasm of all the liver and ductular cells.Binucle-ated,polypoid liver cells and the peripheral cells of the lobules or nodules were distinctly posi-tive,The liver cells which were strongly positive were a kind of thin polygonal cells with asmall oval or a round deeply stained nucleus in each.They might exist sporadically in the lob-ules or in the marginal portion of a nodule.These liver cells are quite different from the so-called oval cells which are derived from the proliferating ductules and are generally believed tobe responsible for the pathogensis of hepatoma.

Antisense oligonucleotide to insulin-like growth factorⅡ induces apoptosis in human ovarian cancer AO cellline

The effects of antisense oligonucleotide to insulin-like growth factor 11 (IGFII) to induce apoptosis in human ovarian cancer cells were evaluated. Antiproliferation effects of antisense to IGFII in ovarian cancer AO cells were determined by 3H-thymidine incorporation. Apoptosis of the IGFll antisense-treated cells was quantitated by both nuclear condensation and flow cytometry after cells were stained with propidium iodide. IGFII antisense (4.5μM)treatment of 48 h maximally inhibited proliferation of AO cells. More than 25% of IGFII antisense-treated cells (4.5PM for 24 h) had undergone apoptosis, whereas less than 3% of the cells were apoptotic in either IGFII sense-treatedcells or untreated cells. Antisense oligonucleotide to IGFII significantly inhibited cell proliferation and induced apoptosis in human ovarian cancer AO cell. These data suggest that IGFII may be a potential target in treatment of ovarian cancer and antisense oligonucleotide to IGFⅡmay serve as a therapeutic approach.

Expression of insulin-like growth factor-1 in breast cancer tissues

Objective: We investigated the expression of insulin-like growth factor-1 (IGF-1) so as to explore its relationship with carcinogenesis and development of breast cancer. Methods: IGF-1 mRNA levels in tissues of breast cancer, adjacent breast cancer in 70 cases breast cancer patients were analyzed by RT-PCR with the normal breast tissues of paired breast as the control. Results: The level of IGF-1 mRNA expression in breast cancer tissues was significantly higher than that in the paired adjacent to breast cancer tissues, normal mammary gland tissues. The ration of IGF-1/β-actin were 0.679 ± 0.075, 0.463 ± 0.085, 0.305 ± 0.031, respectively. There was significant difference between different groups (P < 0.005). Expression of IGF-1 was associated with lymph node metastasis, pathological staging and estrogen receptor status of breast cancer and no significant relationship with tumor pathological grouping (P > 0.005). Conclusion: The high-level expression of IGF-1 in breast cancer tissues is correlated with carcinogenesis, development and metastasis of breast cancer.

The effect of polymorphism in gene of insulin-like growth factor-Ⅰ on the serum periparturient concentration in Holstein dairy cows

Objective:To investigate the relationship between polymorphism within the 5'-untranslated region(5'-UTR)of IGF-I gene and its periparturient concentration in Iranian Holstein dairy cows.Methods:Blood samples(5 mL,n=37)were collected by caudal venipuncture from each animal into sample lubes containing the EDTA and DNA was extracted from blood.In order to measure ICF-I concentration the collection of blood samples(n=111)was also done at 14 d before calving(prepartum),25 and 45 d postpartum.Results:We found evidence for a significant effect of C to T mutation in position 512 of IGF-I gene on its serum concentration in dairy cows in Iran.Cows with CC genotype had significantly higher concentration(Mean-SD)of IGF-I at 14 d prepartum(91.8±18.1)μg/L compared to those with TT genotype(73.3±14.4)μg/L(P=0.04).A significant trend(quadratic)was found for IGF-I concentration,as higher in CC cows compared to ones with TT genotype,during the 14 d before calving to 45 d postpartum(P=0.01).Conclusions:We concluded that C/T transition in the promoter region of IGF-I gene can influence the serum concentration of ICF-I in periparturient dairy cows.

IGF-Ⅱ对肝癌细胞MHCC97-H癌基因C-myc和N-ras表达的影响

目的观察胰岛素样生长因子Ⅱ(IGF-Ⅱ)对肝癌细胞MHCC97-H中癌基因C-myc和N-ras表达的影响。方法培养肝癌细胞株MHCC97-H,用IGF-Ⅱ(50ng/mL)作用48h后,分别采用细胞免疫荧光、Western blot法及Real-time PCR法检测细胞中C-myc和N-ras的表达情况,并与对照组比较,进行定量分析。结果 C-myc及N-ras蛋白阳性表达主要位于细胞核。对照组、IGF-Ⅱ组MHCC97-H细胞中C-myc荧光表达量分别为(100.00±2.89)%、(254.00±35.57)%,N-ras荧光表达量分别为(100.00±14.43)%、(257.30±22.43)%。与对照组相比,IGF-Ⅱ组MHCC97-H细胞中C-myc及N-ras蛋白表达明显增强,差异有统计学意义(P<0.05)。与对照组相比,IGF-Ⅱ组MHCC97-H细胞中C-myc和N-ras的mRNA表达明显增加,差异有统计学意义(P<0.05)。结论 IGF-Ⅱ可上调肝癌细胞MHCC97-H中癌基因C-myc和N-ras的蛋白及mRNA表达,这可能是IGF-Ⅱ促进肝癌细胞增殖的分子机制之一,这为临床上肝癌防治提供了新线索。

IGF-Ⅰ和IGF-Ⅱ在人结直肠腺瘤和结直肠癌中的表达及意义

目的探讨IGF-Ⅰ和IGF-Ⅱ与结直肠腺瘤及结直肠癌发生、发展的关系。方法用免疫组化法检测IGF-Ⅰ和IGF-Ⅱ在人结直肠腺瘤和结直肠癌中的表达,用IPWIN60软件测量染色情况,计算平均光密度。结果 IGF-Ⅰ、IGF-Ⅱ在正常肠粘膜中少量表达,在结直肠管状腺瘤和绒毛状腺瘤中表达明显增强,结直肠癌中则高表达。与正常肠粘膜组比较,结直肠管状腺瘤组、绒毛状腺瘤组、结直肠癌组的表达均有显著差异(P<0.01);与结直肠管状腺瘤组和绒毛状腺瘤组比较,IGF-Ⅰ和IGF-Ⅱ在结直肠癌中的表达显著增强(P<0.01)。结论 IGF-Ⅰ和IGF-Ⅱ在正常结直肠粘膜、管状腺瘤、绒毛状腺瘤、结直肠癌中的表达依次增加,IGF-Ⅰ和IGF-Ⅱ可能在结直肠腺瘤及结直肠癌的发生、发展中起一定作用。

肺癌患者化疗前后血清NSE、IGF-Ⅱ和TNF-α检测的临床意义

目的:探讨肺癌患者化疗前后血清NSE、IGF-Ⅱ和TNF-α含量的变化。方法:应用放射免疫分析检测38例肺癌患者化疗前后血清NSE、IGF-Ⅱ和TNF-α含量,并与35名正常健康人作比较。结果:肺癌患者在化疗前血清中NSE、IGF-Ⅱ和TNF-α水平均非常显著地高于正常人组(P<0.01),化疗后6个月在未复发的25例中明显下降接近于正常人组,而复发的5例,其数值又回升至化疗前水平(P<0.05)。结论:检测肺癌患者血清NSE、IGF-Ⅱ和TNF-α含量的变化可作为诊断和疗效观察的参考。

食管癌患者手术治疗前后血清Hcy、IGF-Ⅱ和TSGF检测的临床意义

目的:探讨了食管癌患者手术治疗前后血清Hcy、IGF-Ⅱ和TSGF水平的变化及意义。方法:应用免疫法和放射免疫分析对33例食管癌患者进行了手术治疗前后血清Hcy、IGF-Ⅱ和TSGF检测,并与35名正常人作比较。结果:食管癌患者在手术前血清Hcy、IGF-Ⅱ和TSGF水平非常显著地高于正常人组(P<0.01)手术后6个月未复发的29例中明显下降接近于正常人组,而复发的4例,其数值又回升至手术前水平(P<0.05)。结论:检测食管癌患者血清Hcy、IGF-Ⅱ和TSGF水平的变化可作为诊断和疗效观察的参考。

IGF-Ⅱ在早期胚胎发育终止蜕膜组织中的表达

目的 研究胰岛素样生长因子 Ⅱ (IGF Ⅱ )在早期胚胎发育终止中的表达以及母体血中IGF Ⅱ水平的改变与胚胎发育终止的关系。方法 采用免疫组织化学SP法检测早期胚胎发育终止蜕膜中IGF Ⅱ蛋白的表达 ;采用原位杂交法检测早期胚胎发育终止蜕膜中IGF ⅡmRNA的表达 ;采用酶联免疫吸附试验 (ELISA)检测正常妊娠及早期胚胎发育终止孕妇血中IGF Ⅱ水平的变化。结果 正常妊娠的蜕膜组织中IGF Ⅱ的表达明显高于胚胎发育终止组 (P<0 .0 1) ;正常妊娠组蜕膜中IGF ⅡmRNA的表达明显高于胚胎发育终止组 (P <0 .0 1) ;正常妊娠组母血中IGF Ⅱ水平也明显高于胚胎发育终止组 (P <0 .0 1)。结论 IGF Ⅱ在蜕膜组织中的表达水平与胚胎发育密切相关 ,其含量的减少可能是导致胚胎死亡的原因之一。

肺癌患者化疗前后血清Hcy、IGF-Ⅱ和TSGF检测的临床意义

目的:探讨肺癌患者化疗前后血清Hcy、IGF-Ⅱ和TSGF水平的变化及临床意义。方法:应用免疫法和放射免疫分析对35例肺癌患者进行了血清Hcy、IGF-Ⅱ和TSGF含量检测,并与30名正常健康人作比较。结果:肺癌患者在化疗前血清Hcy、IGF-Ⅱ和TSGF水平均非常显著地高于正常人组(P<0.01),化疗后6个月在未复发的27例中明显下降接近于正常人组,而复发的8例,其数值又回升至化疗前水平(P<0.05)。结论:检测肺癌患者血清Hcy、IGF-Ⅱ和TSGF水平的变化可作为诊断和疗效观察的参考。

子宫颈癌患者血清IGF-Ⅱ、IL-8和Hcy检测的临床意义

目的:探讨子宫颈癌患者手术治疗前后血清IGF-Ⅱ、IL-8和Hcy检测的临床意义。方法:应用放射免疫分析和免疫化学法对33例子宫颈癌患者进行了手术治疗前后血清IGF-Ⅱ、IL-8和Hcy检测,并与35名正常健康人作比较。结果:子宫颈癌患者在手术治疗前血清IGF-Ⅱ、IL-8和Hcy水平均非常显著地高于正常人组(P<0.01)。手术治疗3个月后则与正常人组比较无显著性差异(P>0.05)。血清IGF-Ⅱ水平与IL-8和Hcy呈显著正相关(r=0.5786、0.6084,P<0.01)。结论:检测子宫颈癌患者手术治疗前后血清IGF-Ⅱ、IL-8和Hcy水平的变化对了解病情、观察疗效和预后判断均具有重要的临床价值。

HDCP患者胎盘组织中抵抗素、IGF-Ⅱ与IGFBP-1的表达及相关性研究

目的探讨妊娠期高血压疾病(HDCP)患者胎盘组织中抵抗素、胰岛素样生长因子-Ⅱ(IGF-Ⅱ)与胰岛素样生长因子结合蛋白-1(IGFBP-1)的表达水平及相关性。方法选取石家庄市第二医院妇产科2015年8月-2018年6月收治的HDCP患者90例作为研究对象,根据诊断分为妊娠期高血压组、轻度子痫前期(MPE)组和重度子痫前期(SPE)组,每组30例。另取30例健康孕妇为对照组。收集分娩后孕妇胎盘组织,免疫组化检测抵抗素、IGF-Ⅱ和IGFBP-1的表达水平,并对结果行相关性分析。结果各组胎盘组织中抵抗素和IGFBP-1水平由高到低依次为SPE组、MPE组、妊娠期高血压组和对照组,组间两两比较差异均有统计学意义(P均<0.05)。各组胎盘组织中IGF-Ⅱ水平由高到低依次为对照组、妊娠期高血压组、MPE组和SPE组,组间两两比较差异均有统计学意义(P均<0.05)。HDCP患者胎盘组织中抵抗素与IGFBP-1水平呈正相关(r=0.356,P<0.05),与IGF-Ⅱ水平呈负相关(r=-0.370,P<0.05);IGFBP-1水平与IGF-Ⅱ水平呈负相关(r=-0.278,P<0.05)。结论与正常孕妇相比,HDCP患者胎盘组织中IGF-Ⅱ水平较低,IGFBP-1和抵抗素水平较高,三者间存在一定相关性,可能参与了HDCP的发病过程。

急性白血病患者化疗前后Hcy、IGF-Ⅱ和TSGF检测的临床意义

目的:探讨了急性白血病患者化疗前后血清Hcy、IGF-Ⅱ和TSGF水平的变化及临床意义。方法:应用免疫法、放免法和生化法对32例急性白血病患者进行了血清Hcy、IGF-Ⅱ和TSGF检测,并与35名正常健康人做比较。结果:急性白血病患者在化疗前血清Hcy、IGF-Ⅱ和TSGF水平非常显著地高于正常人组(P<0.01),化疗后6个月未复发的26例中明显下降接近正常人组,而复发的6例,其数值又回升到化疗前水平(P<0.05)。结论:检测白血病患者血清Hcy、IGF-Ⅱ和TSGF水平的变化可作为诊断和疗效观察的参考指标。

IGF-Ⅱ在肝硬变和肝癌组织的表达及其与LCD的相关性研究

目的 :探讨 IGF- 在肝硬变及肝细胞癌 (HCC)发生发展中的作用及其与不典型增生肝细胞 (L CD)的关系。方法 :运用免疫组织化学 S- P法对 5 7例肝硬变 ,2 9例 HCC癌旁肝组织及 2 1例 HCC进行实验研究。结果 :L CD在肝硬变及癌旁肝组织的发生率分别为 5 6 .1% (32 / 5 7例 )及 6 5 .5 % (19/ 2 9例 )。 IGF- 在 5 7例肝硬变组织、 2 9例癌旁肝组织及 2 1例 HCC中的阳性率分别为 71.9% (41/ 5 7例 ) ,72 .4% (2 1/ 2 9例 )和 47.6 % (10 / 2 1例 ) ,其中伴有 L CD肝组织中 IGF- 阳性率为90 .2 % (46 / 5 1例 ) ,不伴 L CD的肝组织中 IGF- 的阳性率为 45 .7% (16 / 35例 )。结论 :IGF- 的过度表达可能在肝细胞恶性转化早期进程中发挥重要作用 ;L CD可能是肝硬变向 HCC发展过程中的重要中间阶段。

支气管肺炎患儿治疗前后血清IGF-Ⅱ和白三烯测定的临床意义

目的探讨支气管肺炎患儿治疗前后血清IGF-Ⅱ和白三烯水平的变化意义。方法应用放射免疫分析法和酶联法对36例支气管肺炎患儿进行了治疗前后血清IGF-Ⅱ和白三烯水平检测,并与35例正常健康儿作比较。结果在治疗前血清IGF-Ⅱ和白三烯水平非常显著地高于正常人组(P<0.01)。经治疗1周后则与正常人比较差异无显著性(P>0.05)。结论检测支气管肺炎患儿血清中IGF-Ⅱ和白三烯水平对疾病的治疗和预后有十分重要的临床价值。