Effects of miRNA-628-3p on proliferation, apoptosis and invasion of non-small cell lung cancer H1299 cells and its targeting relationship with insulin-like growth factor 1 receptor / 肿瘤研究与临床

Objective:To investigate the effects of miRNA-628-3p (miR-628-3p) on the proliferation, apoptosis and invasion of non-small cell lung cancer H1299 cells and its targeting relationship with insulin-like growth factor 1 receptor (IGF-1R).Methods:The blank control group (untreated H1299 cells), miR-NC group (H1299 cells transfected with empty plasmid), miR-628-3p-M group (H1299 cells transfected with miR-628-3p mimic sequence plasmid) and miR-628-3p-I group (H1299 cells transfected with miR-628-3p inhibitory sequence plasmid) were established. The cells in each group were cultured for 72 h, and the cell proliferation ability was detected by methyl thiazol tetrazolium (MTT) method, the number of cell monoclonal formation was determined by crystal violet staining, the level of cell apoptosis was determined by flow cytometry, and the cell invasion ability was determined by Transwell method. The mRNA levels of miR-628-3p and IGF-1R in cells were determined by real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR), and the protein level of IGF-1R in cells was determined by Western blotting.Results:Compared with the blank control group and miR-NC group, the cell survival rate [(42±7)% vs. (78±6)%, (76±7)%], the number of monoclonal formation [235±35 vs. 614±89, 618±75], the number of invasive cells [(265±85) cells vs. (693±185) cells, (703±119) cells], relative expression of IGF-1R mRNA (2.17±0.14 vs. 3.38±0.15, 3.37±0.13) and relative expression of IGF-1R protein (0.34±0.13 vs. 0.89±0.19, 0.88±0.18) in the miR-628-3p-M group were lower (all P < 0.05), but the apoptosis rate [(9.30±3.51)% vs. (3.30±1.54)%, (3.10±1.94)%] and relative expression of miR-628-3p (6.93±0.17 vs. 3.29±0.15, 3.30±0.16) were higher (all P < 0.05); the cell survival rate [(90±6)%], the number of monoclonal formation (1 063±102), the number of invasive cells [(1 985±426) cells], relative expression of IGF-1R mRNA (4.30±0.18) and relative expression of IGF-1R protein (1.47±0.17) in the miR-628-3p-I group were higher (all P < 0.05), but the apoptosis rate [(0.90±0.20)%] and the relative expression of miR-628-3p (1.93±0.18) were lower (both P < 0.05). Compared with the miR-628-3p-M group, the miR-628-3p-I group had higher cell survival rate, the number of monoclonal formation, the number of invasive cells, and the relative expressions of IGF-1R mRNA and protein (all P < 0.05), but the apoptosis rate and relative expression of miR-628-3p were lower (both P < 0.05). Conclusions:After regulation of miR-628-3p level, the proliferation, migration and invasion of H1299 cells are affected. miR-628-3p may have a targeting relationship with IGF-1R.

Insulin-like growth factor-1 receptor inhibitor alleviates diabetic kidney disease mouse tubulopathy / 中华肾脏病杂志

Objective@#To investigate the effects of insulin-like growth factor 1 receptor (IGF-1R) inhibitor on tubulopathy in diabetic kidney disease (DKD) mice.@*Methods@#C57BL/6J male mice were randomly divided into normal control group (n=10) and DKD model group (n=30), by giving a single intraperitoneal injection of STZ 150 mg/kg to establish a DKD model. After established successfully, the mice in DKD model group were randomly divided into DKD group (n=10), benazepril group (n=10) and IGF-1R inhibitor group (n=10). IGF-1R inhibitor group was given intraperitoneal injection of IGF-1R inhibitor (30 mg·kg-1·d-1) and benazepril group was given intraperitoneal injection of benazepril (30 mg·kg-1·d-1). Normal control group and DKD group were given an equal amount of normal saline. After 8 weeks of feeding, mice were euthanatized. Body weight and kidney weight were recorded. Blood, urine and kidney samples were collected. Biochemical tests such as blood glucose and urine albumin were measured by automatic biochemical instruments and albumin excretion rate was calculated. Pathological changes of mice were observed by hematoxylin-eosin staining (HE) and periodic acid-schiff staining (PAS). Phosph (p) IGF-1R expression level was determined by immunohistochemistry and Western blotting.@*Results@#Compared with the normal control group, blood glucose, kidney weight/body weight and urinary albumin excretion rate were significantly higher in DKD group (all P<0.01). In DKD mice, glomerular expansion, tubular stenosis, tubular swelling and tubular atrophy were significantly detected. Meanwhile, the number of proximal tubular epithelial (PTE) cells was decreased, and the renal tubular injury scores, the average glomerular volume, and pIGF-1R protein expression were increased (all P<0.05). Compared with the DKD group, albumin excretion rate was significantly reduced (P<0.01), the above pathological changes were alleviated and the effect of IGF-1R inhibitor was more significant. Compared with the DKD group, the pIGF-1R protein expression was reduced in IGF-1R inhibitor group (P<0.05). Compared with the benazepril group, the pIGF-1R protein expression was reduced in IGF-1R inhibitor group (P<0.05).@*Conclusion@#IGF-1R inhibitor has better effect than benazepril on alleviating the tubulopathy of DKD mice.

Insulin-like growth factor-1 receptor inhibitor alleviates diabetic kidney disease mouse tubulopathy / 中华肾脏病杂志

Objective To investigate the effects of insulin-like growth factor 1 receptor (IGF-1R) inhibitor on tubulopathy in diabetic kidney disease (DKD) mice.Methods C57BL/6J male mice were randomly divided into normal control group (n=10) and DKD model group (n=30),by giving a single intraperitoneal injection of STZ 150 mg/kg to establish a DKD model.After established successfully,the mice in DKD model group were randomly divided into DKD group (n=10),benazepril group (n=10) and IGF-1R inhibitor group (n=10).IGF-1R inhibitor group was given intraperitoneal injection of IGF-1R inhibitor (30 mg· kg-1· d-1) and benazepril group was given intraperitoneal injection of benazepril (30 mg· kg-1· d-1).Normal control group and DKD group were given an equal amount of normal saline.After 8 weeks of feeding,mice were euthanatized.Body weight and kidney weight were recorded.Blood,urine and kidney samples were collected.Biochemical tests such as blood glucose and urine albumin were measured by automatic biochemical instruments and albumin excretion rate was calculated.Pathological changes of mice were observed by hematoxylin-eosin staining (HE) and periodic acid-schiff staining (PAS).Phosph (p) IGF-1R expression level was determined by immunohistochemistry and Western blotting.Results Compared with the normal control group,blood glucose,kidney weight/body weight and urinary albumin excretion rate were significantly higher in DKD group (all P ＜ 0.01).In DKD mice,glomerular expansion,tubular stenosis,tubular swelling and tubular atrophy were significantly detected.Meanwhile,the number of proximal tubular epithelial (PTE) cells was decreased,and the renal tubular injury scores,the average glomerular volume,and plGF-1R protein expression were increased (all P ＜ 0.05).Compared with the DKD group,albumin excretion rate was significantly reduced (P ＜ 0.01),the above pathological changes were alleviated and the effect of IGF-1R inhibitor was more significant.Compared with the DKD group,the pIGF-1R protein expression was reduced in IGF-1R inhibitor group (P ＜ 0.05).Compared with the benazepril group,the pIGF-1R protein expression was reduced in IGF-1R inhibitor group (P ＜ 0.05).Conclusion IGF-1R inhibitor has better effect than benazepril on alleviating the tubulopathy of DKD mice.

Expressions and Clinical Significance of Insulin-like Growth Factor-1 and its Receptor in Patients With Type 2 Diabetes Mellitus and Gastric Cancer / 胃肠病学

Insulin-like growth factor-1 (IGF-1) is an important mitogenic factor, which has an intense role in promoting proliferation and anti-apoptosis, and is abnormally expressed in many malignant tumors. Aims: To investigate the expressions and clinical significance of IGF-1 and its receptor (IGF-1R) in patients with type 2 diabetes mellitus (T2DM) and gastric cancer (GC). Methods: A total of 338 patients with GC received surgery from Jan. 2010 to Jan. 2013 were enrolled, and were divided into GC group and GC-T2DM group. The expressions of IGF-1 and IGF-1R were detected by immunohistochemistry, and their relationship with clinicopathological parameters were analyzed. The effect of expressions of IGF-1 and IGF-1R on survival of patients was evaluated. Results: Compared with GC group, the positivity rates of IGF-1 (81.3% vs. 42.3%; χ2=31.427, P0.05). Conclusions: Compared with GC patients, the positivity rates of IGF-1 and IGF-1R in GC-T2DM patients are significantly increased, and have certain negative effects on the clinicopathological parameters of the patients, suggesting that the two may promote the development and progression of GC through some mechanism.

The effect of levonorgestrel-releasing intrauterine system on IGF-Ⅰ and IGF-IR expression after transcervical resection of polyp / 中国医师杂志

Objective To evaluate the effect of levonorgestrel-releasing intrauterine system (LNG-IUS) on insulin-like growth factor-Ⅰ (IGF-Ⅰ) and insulin-like growth factor-Ⅰ receptor (IGF-IR) expression after transcervical resection of polyp (TCRP).Methods 100 cases of endometrial polyps were selected.The control group (n =50) was treated with TCRP only,while the observation group (n =50) was treated with LNG-IUS after TCRP.The scores of pictorial blood loss assessment chart (PBCA),endometrial thickness,recurrence rate,mRNA expression of IGF-Ⅰ and IGF-IR in endometrial tissue were compared between the two groups.Results At 1,3,6,12 months follow-up,the PBAC score and endometrial thickness of observation group were significantly lower than control group (P ≤ 0.05).At 12 months after operation,the mRNA expression levels of IGF-Ⅰ and IGF-IR in the endometrium of the observation group were significantly lower than those of the control group (P ≤ 0.05).After 12 months of follow-up,the recurrence rates of the control group and the observation group were 16.0％ (8/50) and 4.0％ (2/50),respectively.The recurrence rate of the observation group was significantly lower than that of the control group (P ≤ 0.05).Conclusions TCRP combined with LNG-IUS treatment can significantly reduce EP recurrence,and down-regulation of the mRNA expression of IGF-Ⅰ and IGF-IR maybe its possible mechanism.

Clinicopathological and prognostic analysis of insulin like growth factor 1 receptor protein expression in primary gastrointestinal stromal tumors / 中华普通外科杂志

Objective To investigate the clinicopathological significance of expression of insulin like growth factor 1 receptor (IGF1R) protein in primary gastrointestinal stromal tumors (GIST) and their impacts on prognosis.Methods Between January,2005 and January,2011,84 primary GIST patients underwent surgery.Immunohistochemical analysis was performed based on tissue microarray to estimate expression pattern of IGF1R in tumor cells and nomal controls.Association of IGF1R expression with clinicopathological features and relapse-free survival (RFS) were also analyzed.Results The negative,weakly positive,positive,and strong positive expression rates of IGF1 R protein in GIST group were 20％,14％,48％ and 18％,respectively;while in the control group were 32％,40％,20％ and 8％,respectively (x2 =30.663,P ＜ 0.001).The 5-year overall survival (OS) rate was 93％.1-year,3-year and 5-year RFS rate were 99％,76％ and 60％,respectively.As shown by univariate analysis the following factors were poor prognostic indicators for RFS,non-gastric tumor location (P =0.017),large tumor size (P =0.022),high mitotic index (P ＜ 0.001),high cellularity (P =0.012),tumor rupture (P =0.013),absent or low expression of IGF1R (P =0.022).Tumor size (HR5.1-10 ≤5 cm =1.86,95％ CI:0.67-5.15;HR＞10 ≤5cm =6.71,95％ CI:0.67-5.15,overall P =0.023),and mitotic index (HR5.1-10/50HPFsvs.≤5/50HPFs =5.72,95％ CI:2.09-15.64;HR＞10/50HPF ≤5/50HPFs =3.44,95％ CI:1.13-10.45,overall P =0.002) were negative independent risk predictors by multivariate analysis.Conclusions High expression of IGF1R may be involved in the occurrence,development and poor prognostic of primary GIST.Expression of IGF1 R is correlated with high risk potential and may predict early recurrence.

Research advances in insulin-like growth factor 1 receptor and its pathway in diagnosis and treatment of primary liver cancer / 临床肝胆病杂志

The main biological functions of insulin-like growth factor 1 receptor (IGF-1 R) include formation and maintenance of transformed cell phenotype,involvement in cell proliferation and differentiation,and inhibition of cell apoptosis.In addition,IGF-1R regulates cell cycle and works with epidermal growth factor and platelet-derived growth factor to mediate cells to enter S phase from G1 phase.Overexpressed IGF-1R has become one of the target proteins for diagnostic imaging and localization therapy for primary liver cancer.Inhibition of the expression or function of IGF-1R can effectively control the growth and metastasis of tumor cells and enhance their sensitivity to chemotherapy and radiotherapy.This article reviews the role and significance of IGF-1R and its pathway in the diagnosis and treatment of primary liver cancer.

Research advances in insulin-like growth factor-1 receptor in pancreatic cancer / 临床肝胆病杂志

Insulin-like growth factor-1 receptor (IGF-1 R) widely exists in the surface of various types of cells and is closely associated with the formation and development of tumor cells.It also provides a new direction for the targeted therapy for tumors.This article reviews the expression,development,and progression of IGF-1R in pancreatic cancer and research advances in IGF-1R as a target for tumor treatment.

miR-126-mediated activation of IGF2/IGF1R/IRS1 signaling promotes the Herceptin resistance in ErbB2 positive breast cancer cells / 中国医师杂志

Objective To explore the role of insulin-like growth factor-2/insulin-like growth factor1 receptor/insulin receptor substrate-1 (IGF2/IGF1R/IRS1) signal pathway inducing the chemoresistance of epidermal growth factor receptor 2 (ErbB2) positive breast cancer cells to Herceptin.Methods Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assay were used to determine the expression levels of IGF2,IGF1 R,and IRS1.The direct targets of miR-126 were validated by dual-luciferase reporter gene assay.In SKBR3/pool2 cells,IGF1 R activity was reduced by an inhibitor of IGF1 R,and IRS1 was knocked-down by shRNAs.Furthermore,3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to evaluate the sensitivity of these treated cells to Herceptin.Results IGF2,IGF1 R,and IRS1 were significantly higher expressed in SKBR3/pool2 cell compared to that in SKBR3 cell.Western blot assay showed that IGF2/IGF1R/IRS1 was activated in SKBR3/pool2 cells.Bioinformatics analysis combined with luciferase activity suggested that miR-126 directly targeted IRS1.MTS results demonstrated that the chemosensitivity to Herceptin of SKBR3/ pool2 cells with inhibitor of IGF1R or shRNAs targeting IRS1 or overexpressing miR-126 was significantly reduced.Conclusions IGF2/IGF1R/IRS1 signal pathway confers to the chemoresistance of ErbB2 positive breast cancer cells to Herceptin.

The function and mechanism of ErbB3 and IGF1R in breast cancer with Herceptin resistance / 中国医师杂志

Objective To investigate the role of epidermal growth factor receptor 3 (ErbB3) and insulin-like growth factor-1 receptor (IGF1R) in enhancing the resistance of Herceptin in human breast cancer.Methods HRG (Heregulin,the ligand of ErbB3) or IGF2 (insulin-like growth factor2,the ligand of IGF1R) was correspondingly added into breast cancer cells SKBR3 and BT474,and then 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and were performed in these cells to evaluate the sensitivity of these cells to Herceptin.Furthermore,we used HRG or IGF2 antibodies to inhibit their joint receptors in Herceptin-resistant breast cancer cells SKBR3/POOL2 and BT474/HR20.Finally,the sensitivity of these treated cells to Herceptin was detected via MTS assay.HRG or IGF2 was added into breast cancer cell BT474,and co-IP assay was used to detect the expressions of ErbB3 and IGF1R which combined with ErbB2.Results The treatment groups used HRG or IGF2 enhanced the resistance of Herceptin in Herceptin-sensitive breast cancer cells.On the other hand,we used antibodies of HRG and IGF2 to block their combining with their receptors in Herceptin-resistant breast cancer cells,the cells became more sensitive to Herceptin.BT474 cell was treated with HRG or IGF2.The expressions of ErbB3 and IGF1R which combined with ErbB2 were increased.Conclusions The formation of heterodimers ErbB2/ErbB3 and ErbB2/IGF1R might enhance the resistance of Herceptin in ErbB2-overexpression human breast cancers.

Relation between cervical squamous cancer lesion among Uighur and Han ethnic women in Xinjiang with expression of PI3K,ERK,IGF-1R and ER protein / 重庆医学

Objective To observe the expression levels of PI3K ,ERK ,IGF‐1R and ER in cervicitis and cervical squamous cancer tissues among Uighur and Han ethnic patients and their correlation .Methods The 90 paraffin embedding samples of cervici‐tis tissue( 46 cases for Han and 44 cases for Uighur) and 224 paraffin embedding samples of cervical squamous cancer tissue (36 ca‐ses for Han and 188 cases for Uighur) were collected and detected the protein expression levels by using immunohistochemistry .Re‐sults The positive expression rates of IGF‐1R and PI3K in cervical squamous cancer were 58 .04% and 92 .41% respectively ,which were higher than 13 .33% and 57 .78% in cervicitis tissue ,the positive expression rates of ER and ERK in cervical squamous cancer were 22 .32% and 68 .30% respectively ,which were lower than 63 .33% and 95 .56% in cervicitis tissue ;the positive expression rate of IGF‐1R and PI3K of cervical squamous cancer in Han and Uighur were 69 .44% ,88 .89% and 55 .85% ,93 .09% respective‐ly ,which were higher than 15 .22% ,54 .35% and 11 .36% ,61 .36% of cervicitis tissue ;the positive expression rate of ER and ERK of cervical squamous cancer in Han and Uighur were 13 .89% ,83 .33% and 23 .94% ,65 .43% respectively ,which were lower than 65 .22% ,93 .48% and 61 .36% ,97 .73% of cervicitis tissue respectively ;the expression of ERK in Uighur cervical squamous carci‐noma tissue was 65 .43% ,which was lower than 83 .33% in Han ,the difference was statistically significant (P<0 .01) .Conclusion PI3K ,ERK ,IGF‐1R and ER protein expression positive or deficiency is closely related to the occurrence of cervical cancer ,which may serve as the important biological indicators for detecting cervical cancer ,and the ethnic difference of ERK protein expression exists in cervical cancer .

Expressions of IGF-1R and IGFBP-3 in colon cancer metastasis and their correlations with lymphatic metastasis / 国际肿瘤学杂志

Objective To investigate the expressions of insulin-like growth factor receptor-1 (IGF-1R) and insulin-like growth factor binding protein-3 (1GFBP-3) and their correlations with clinicopathological parameters in the primary colon cancer,as well as their roles in lymph node metastasis of colon cancer.Methods The expressions of IGF-1R and IGFBP-3 in 78 cases of colon cancer tissues and 78 cases of normal colon mucosa tissues were detected by SP immunohistochemical technology and the correlations between the expressions and the clinical pathological parameters of colon cancer were analyzed.Results The positive rate of IGF-1R in colon cancer (66.7％,52/78) was significantly higher than that in control group (24.4％,19/78),x2 =28.150,P =0.000.The positive rate of IGFBP-3 in colon cancer (73.1％,57/78) was significantly lower than that in control group (89.7％,70/78),x2 =7.158,P =0.007.IGF-1R expression in colon cancer was significantly correlated with the invasion (x2 =5.804,P =0.016),TNM stage (x2 =5.246,P =0.022) and lymph node metastasis (x2 =12.955,P =0.000).IGFBP-3 expression in colon cancer was signi-ficantly correlated with the TNM stage (x2 =7.096,P=0.008),lymph node metastasis (x2 =5.893,P =0.015) and distant metastasis (P =0.003).Both with other factors had no significant correlation (P ＞ 0.05).1GF-1R expression and IGFBP-3 expression showed a negative correlation (r =-0.245,P =0.03).Conclusion The over expression of IGF-1R and low expression of IGFBP-3 are associated with TNM stage and lymph node metastasis in colon cancer.IGF-1R and IGFBP-3 may become new targets of the treatment of colon cancer.

Effects of hyperbaric oxygen on the expressions of hypoxia-inducible factor 1α and insulin-like growth factor-1 receptor in the formation of hyperplastic scar in rabbit ears / 中华皮肤科杂志

Objective To evaluate the effects of hyperbaric oxygen on the expressions of hypoxia-inducible factor 1α (HIF-1α) and insulin-like growth factor-1 receptor (IGF-1R) in the formation of hyperplastic scar in rabbit ears.Methods The ears of 20 New Zealand rabbits were used to construct an animal model for hyperplastic scar by operation.After the establishment of scar models,the rabbits were randomly divided into 4 experimental groups and one control group with 4 mice (48 wound surfaces) in each group.The mice in the 4 experimental groups were treated with hyperbaric oxygen for 7,14,21 and 28 days,respectively,and those in the control group remained in normoxic environment after operation.Scar tissues were resected from all the rabbit ears on day 29 after operation.Hematoxylin and eosin (HE) staining was conducted for the observation of morphological changes and calculation of scar elevation index,and immunohistochemistry to measure the expressions of HIF-1α and IGF-1R.Statistical analysis was carried out by one-way analysis of variance followed by least significant difference t-test.Results HE staining showed that both the number of fibroblasts and amount of collagen fibers were significantly reduced in the experimental groups compared with the control group.Scar elevation index was 4.28 ± 0.22 in the control group,3.64 ± 0.29,3.46 ± 0.21,3.29 ± 0.21,3.16 ± 0.15 in the 7-,14-,21-and 28-day experimental groups respectively,with significant differences among these groups (F =77.70,P ＜ 0.05).The expressions of HIF-1α and IGF-1R were significantly lower in these experimental groups than in the control group (all P ＜ 0.01),lower in the 14-day group than in the 7-day group (P ＜ 0.05),and lower in the 21-day group than in the 14-day group (P ＜ 0.05),with no significant differences between the 28-day group and 21-day group (both P ＞ 0.05).Conclusion Hyperbaric oxygen can effectively down-regulate the expressions of HIF-1α and IGF-1R in scar tissue,and significantly inhibit the formation of hyperplastic scar in rabbit ears.

Análise da expressão e do silenciamento do receptor tipo 1 do fator de crescimento semelhante à insulina em tumores adrenocorticais humanos / Analysis of expression and silencing of insulin-like growth factor 1 receptor in human adrenocortical tumors

Introdução O sistema dos fatores de crescimento semelhantes à insulina (IGF) desempenha importante papel no crescimento e desenvolvimento celular normal. Hiperexpressão do gene IGF1R tem sido demonstrada em diversos tumores, sugerindo que a expressão deste receptor represente um pré-requisito fundamental para transformação celular. Nosso grupo de pesquisa demonstrou o aumento de expressão de IGF1R em tumores adrenocorticais pediátricos. Objetivos: Induzir o silenciamento do gene IGF1R por siRNA na linhagem de tumor adrenocortical humano NCI H295R, bem como avaliar os efeitos in vitro por meio da análise de proliferação celular e apoptose desta linhagem celular. Adicionalmente, avaliar a expressão de IGF-1R e de microRNAs relacionados a sua transcrição em tumores adrenocorticais humanos. Pacientes e métodos: A linhagem celular de carcinoma adrenocortical humano NCI H295R foi cultivada e submetida ao tratamento com 2 siRNAs específicos para IGF-1R. Todos os experimentos foram realizados em quatro grupos: (1) células não tratadas com siRNA, (2) células tratadas com siRNA # 1, (3) células tratadas com siRNA # 2 e (4) células tratadas com o siRNA controle negativo. A expressão gênica e proteica de IGF-1R foram determinadas por meio das técnicas de PCR em tempo real e Western Blot, respectivamente. Os efeitos do silenciamento de IGF-1R in vitro foram avaliados por ensaios de proliferação celular e análise de atividade de caspases. Além disso, 202 pacientes com tumor adrenocortical foram selecionados para o estudo de expressão proteica de IGF-1R por imunohistoquímica. Para avaliação de expressão de microRNAs relacionados à expressão de IGF-1R (miR-100, 375, 145 e 126) por PCR em tempo real foram selecionados 32 pacientes dos 202 disponíveis. Resultados: A expressão de IGF-1R foi significantemente diminuída nas células tratadas com siRNA # 1 e siRNA # 2. Os valores relativos de RNA mensageiro de IGF1R diminuíram aproximadamente 50% e as análises de Western Blot...

Introduction: The insulin-like growth factor (IGF) system plays a key role in normal cell growth and development. IGF1R overexpression has been demonstrated in several tumors suggesting that its expression is a prerequisite for cell transformation. We demonstrated IGF1R overexpression in pediatric adrenocortical tumors. Objectives: To induce IGF1R silencing by siRNA in a human adrenocortical cell line NCI H295R and evaluate its effects on cell proliferation and apoptosis. Additionally, evaluate the expression of IGF-1R protein and microRNAs related to its transcription in human adrenocortical tumors. Patients and methods: The human adrenocortical tumor cell line NCI H295R was cultured and treated with 2 specific IGF1R siRNA. All experiments were carried out in four groups: (1) untreated NCI H295R cells, (2) NCI H295R cells transfected with specific IGF1R siRNA # 1, (3) NCI H295R cells transfected with specific IGF1R siRNA # 2 and (4) NCI H295R cells transfected with a negative control. IGF-1R gene and protein expression was determined by the techniques of real-time PCR and Western blot, respectively. We assessed the effects of IGF-1R silencing on cell proliferation and apoptosis. Moreover, 202 patients with adrenocortical tumors were selected for the study of IGF-1R protein expression by immunohistochemistry. In the analysis of microRNAs that are related to IGF1R (miR-100, 375, 145 e 126) by real time PCR, 32 out 202 patients were selected. Results: IGF-1R levels were significantly decreased in cells that were treated with IGF-1R siRNA # 1 and siRNA # 2. The relative values of IGF1R mRNA decreased approximately 50% and Western blot analysis revealed a 30% of reduction in IGF-1R protein. Downregulation of this gene was accompanied by a reduction in 40% of cell growth in vitro and an increase in 45% of apoptosis. The analysis of microRNAs demonstrated that IGF1R expression is not correlated with the expression of these small RNAs. Additionally, the analysis of...

Growth Hormone-Releaser Diet Attenuates Cognitive Dysfunction in Klotho Mutant Mice via Insulin-Like Growth Factor-1 Receptor Activation in a Genetic Aging Model

BACKGROUND: It has been recognized that a defect in klotho gene expression accelerates the degeneration of multiple age-sensitive traits. Accumulating evidence indicates that aging is associated with declines in cognitive function and the activity of growth hormone (GH)/insulin-like growth factor-1 (IGF-1). METHODS: In this study, we examined whether a GH-releaser diet could be effective in protecting against cognitive impairment in klotho mutant mice. RESULTS: The GH-releaser diet significantly induced the expression of IGF-1 and IGF-1 receptors in the hippocampus of klotho mutant mice. Klotho mutant mice showed significant memory impairments as compared with wild-type mice. In addition, the klotho mutation significantly decreased the expression of cell survival/antiapoptotic factors, including phospho-Akt (p-Akt)/phospho-glycogen synthase kinase3beta (p-GSK3beta), phospho-extracellular signal-related kinase (p-ERK), and Bcl-2, but significantly increased those of cell death/proapoptotic factors, such as phospho-c-jun N-terminal kinase (p-JNK), Bax, and cleaved caspase-3 in the hippocampus. Treatment with GH-releaser diet significantly attenuated both decreases in the expression of cell survival/antiapoptotic factors and increases in the expression of cell death/proapoptotic factors in the hippocampus of klotho mutant mice. In addition, klotho mutation-induced oxidative stress was significantly attenuated by the GH-releaser diet. Consequently, a GH-releaser diet significantly improved memory function in the klotho mutant mice. GH-releaser diet-mediated actions were significantly reversed by JB-1, an IGF-1 receptor antagonist. CONCLUSION: The results suggest that a GH-releaser diet attenuates oxidative stress, proapoptotic changes and consequent dysfunction in klotho mutant mice by promoting IGF-1 expression and IGF-1 receptor activation.

Prognostic Significance of p53, mTOR, c-Met, IGF-1R, and HSP70 Overexpression after the Resection of Hepatocellular Carcinoma

BACKGROUND/AIMS: The current study examines the expression of molecular biomarkers in hepatocellular carcinoma (HCC) and whether these findings correlate with the clinicopathologic features of the disease and patient survival. METHODS: We analyzed the immunohistochemical expression of p53, mammalian target of rapamycin (mTOR), c-Met, and insulin-like growth factor 1 receptor (IGF-1R) heat shock protein 70 (HSP70) with the clinicopathologic features of 83 HCCs. RESULTS: p53 expression was higher in the male patients with undifferentiated histological tumor grades, cirrhosis, and portal vein invasion. High 48 c-Met expression correlated with cirrhosis, and high mTOR expression correlated with the tumor grade and cirrhosis. High IGF-1R expression correlated with the tumor grade and cirrhosis. A multivariate analysis identified a significant relationship between the high expression of p53, tumor grade, and portal vein invasion. In addition, a high expression of mTOR was related to tumor grade and cirrhosis, and a high expression of HSP70 was related to portal vein invasion in a multivariate analysis. The Kaplan-Meier survival curve for patients with high versus low Edmondson grades and p53 expression was statistically significant. CONCLUSIONS: p53, mTOR, and IGF-1R expression correlated with the Edmondson tumor grade in a univariate analysis, while p53 and mTOR correlated with the Edmondson tumor grade in a multivariate analysis. In addition, the tumor grade was found to predict survival. p53 was primarily related to the clinicopathologic features compared to other markers, and it is a poor prognostic factor of survival.

A study on CD90, IGF1R expression in hepatocellular carcinoma / 中华普通外科杂志

Objective To investigate the expression of CD90,IGF1R in hepatocellular carcinoma.Methods CD90,IGF1R expression were detected by SP immunohistochemical staining in 36 cases of hepatocellular carcinoma,20 cases of normal liver tissue biopsied from patients of chronic cholecystitis undergoing cholecystectomy. Results The positive rate of CD90 (63.89％,23/36),IGF1R (52.78％,19/36) in hepatocellular carcinoma significantly increased (P ＜ 0.05 ); The positive rate of CD90 was higher in UICC Ⅲ - Ⅳ stage group (79.17％ ) than in UICC stage Ⅰ - Ⅱ group (33.33％,P ＜0.05) ;The positive rate of CD90 was higher in low differentiated group (76.92％ ) than in well-differentiated group (56.52％,P ＜0.05).The positive rate of IGF1R was higher in UICC Ⅲ - Ⅳ stage group (70.83％ ) than in UICC stage Ⅰ - Ⅱ group ( 16.67％,P ＜ 0.05 ). The positive rate of IGF1R was higher in low differentiated group (84.62％) than in well-differentiated group (37.38％,P ＜ 0.05 ).The expression of IGF1R was positively correlated with that of CD90 (P ＜ 0.05 ).The median survival of CD90+ patients (85 days) was shorter than CD90- patients ( 505 days ) ( P ＜ 0.05 ).The median survival of IGF1 R + patients (100 days) was shorter than IGF1 R- patients (408 days,P ＜ 0.05). Conclusions CD90 or IGF1R expression correlates positively with the progression of HCC.

Mutações no gene do receptor do fator de crescimento insulina-símile 1 (IGF1R) como causa de retardo do crescimento pré- e pós-natal / Mutations in insulin-like growth factor receptor 1 gene (IGF1R) resulting in intrauterine and postnatal growth retardation

Aproximadamente 10 por cento das crianças nascidas pequenas para a idade gestacional (PIGs) não apresentam recuperação espontânea do crescimento. As causas desse déficit de crescimento pré-natal e sua manutenção após o nascimento ainda não são completamente conhecidas na maioria dos casos. Nos últimos oito anos, diversas mutações inativadoras e deleções do gene IGF1R em heterozigose foram relatadas, indicando o papel de defeitos no eixo IGFs/IGF1R como causa do déficit de crescimento. Postula-se que pelo menos 2,5 por cento das crianças nascidas PIGs possam apresentar defeitos no gene IGF1R. O quadro clínico desses pacientes apresenta grande variabilidade quanto à gravidade do retardo de crescimento e aos parâmetros hormonais. Nos casos mais evidentes, os pacientes apresentam microcefalia, déficit cognitivo leve e valores elevados de IGF-1, associados à baixa estatura de início pré-natal. Esta revisão abordará os aspectos clínicos, moleculares e do tratamento da baixa estatura com hrGH de crianças com mutações no IGF1R.

Approximately 10 percent of children born small-for-gestational age (SGA) do not show spontaneous growth catch-up. The causes of this deficit in prenatal growth and its maintenance after birth are not completely known, in most cases. Over the past eight years, several heterozygous inactivating mutations and deletions in IGF1R gene have been reported, indicating the role of defects in the IGFs/IGF1R axis as a cause of growth deficit. It has been hypothesized that at least 2.5 percent of children born SGA may have IGF1R gene defects. The clinical presentation of these patients is highly variable in the severity of growth retardation and hormonal parameters. In the most evident cases, patients have microcephaly, mild cognitive impairment and high levels of IGF-1, associated with short stature of prenatal onset. This review will describe the clinical, molecular and treatment of short stature with hrGH of children with mutations in the IGF1R gene.

A combination with IGF-1R inhibitior enhances the sensitivity of human non-small cell lung cancer cell line PC9/G to gefitinib / 肿瘤

Objective: To observe the effect of gefitinib alone or in combination with AG1024 (a tyrosine kinase inhibitor of insulin-like growth factor-1 receptor) on drug resistance of human non-small cell lung cancer cell line PC9/G with acquired-resistance to gefitinib, and to discuss its possible mechanism. Methods: PC9/G cells were treated with AG1024 or gefitinib alone or in combination, the proliferative activity of PC9/G cells was detected by MTT method, and the efficacy of combination therapy was assessed by median-effects principle. Apoptosis rate of PC9/G cells was examined by flow cytometry. The expression levels of phosphorylated epidermal growth factor receptor (p-EGFR), phosphorylated-Akt (p-Akt) and the phosphorylated extracellular signal-regulated kinase (p-ERK) in PC9/G cells were detected by Western blotting. Results: PC9/G cells displayed apoptotic features after treatment with AG1024 or gefitinib alone, and the cell proliferation was inhibited to different degrees. The treatment of AG1024 combined with gefitinib had a synergistic effect on the apoptosis and the cell proliferation (P<0.05) of PC9/G cells, as well as the expression levels of p-EGFR, p-Akt and p-ERK proteins were decreased. Conclusion: AG1024 in combination with gefitinib exerts a synergistic effect, which may lead to the proliferation inhibition and the apoptosis enhancement, and eventually increases the sensitivity of human non-small cell lung cancer cell line PC9/G to gefitinib. Copyright© 2011 by the Editorial Board of Tumor.