Lenvatinib down-regulates IGF1R/Mek/Erk signaling pathway in the treatment of regorafenib-resistant hepatocellular carcinoma / 中华肿瘤杂志

Objective: To investigate the therapeutic effect and mechanism of lenvatinib on regorafenib-resistant hepatocellular carcinoma cells. Methods: CCK-8 and clone formation assay were used to observe the inhibitory effect of lenvatinib on the growth of hepatocellular carcinoma cells. Flow cytometry was used to detect the apoptosis of regorafenib-resistant hepatocellular carcinoma cells treated with lenvatinib. The expression levels of related proteins were detected by western blot and immunohistochemical staining. The inhibitory effect of lenvatinib on the tumor formation ability of regorafenib-resistant hepatocellular carcinoma cells in vivo was observed by subcutaneous tumor formation experiment in mice. Results: CCK-8 and clone formation assay showed that lenvatinib could inhibit the proliferation of regorafenib-resistant hepatocellular carcinoma cells. The number of clones of HepG2, SMMC7721 and regorafenib-resistant HepG2, SMMC7721 cells in lenvatinib group (120.67±11.06, 53.00±11.14, 55.00±9.54, 78.67±14.64) were all lower than those in control group (478.00±24.52, 566.00±27.87, 333.67±7.02, 210.00±12.77, all P<0.05). Flow cytometry showed that lenvatinib could promote apoptosis of regorafenib-resistant hepatocellular carcinoma cells, the apoptosis rates of HepG2, SMMC7721 and regorafenib-resistant HepG2, SMMC7721 cells in lenvatinib group [(12.30±0.70)%, (9.83±0.38)%, (15.90±1.32)%, (10.60±0.00)%] were all higher than those in control group [(7.50±0.87)%, (5.00±1.21)%, (8.10±1.61)%, (7.05±0.78)%, all P<0.05]. The apoptosis-related protein levels suggested that apoptosis was increased in the treatment of lenvatinib. The animal study showed that lenvatinib can inhibit the growth of regorafenib-resistant cells in vivo. Immunohistochemistry and western blot results showed that lenvatinib could down-regulate the abnormally activated IGF1R/Mek/Erk signaling pathway in regorafenib-resistant cells. Conclusion: Lenvatinib can reverse regorafenib resistance in hepatocellular carcinoma, possibly by down-regulating IGF1R/Mek/Erk signaling pathway.

Translocation of IGF-1R in endoplasmic reticulum enhances SERCA2 activity to trigger Ca2+ER perturbation in hepatocellular carcinoma

The well-known insulin-like growth factor 1 (IGF1)/IGF-1 receptor (IGF-1R) signaling pathway is overexpressed in many tumors, and is thus an attractive target for cancer treatment. However, results have often been disappointing due to crosstalk with other signals. Here, we report that IGF-1R signaling stimulates the growth of hepatocellular carcinoma (HCC) cells through the translocation of IGF-1R into the ER to enhance sarco-endoplasmic reticulum calcium ATPase 2 (SERCA2) activity. In response to ligand binding, IGF-1Rβ is translocated into the ER by β-arrestin2 (β-arr2). Mass spectrometry analysis identified SERCA2 as a target of ER IGF-1Rβ. SERCA2 activity is heavily dependent on the increase in ER IGF-1Rβ levels. ER IGF-1Rβ phosphorylates SERCA2 on Tyr990 to enhance its activity. Mutation of SERCA2-Tyr990 disrupted the interaction of ER IGF-1Rβ with SERCA2, and therefore ER IGF-1Rβ failed to promote SERCA2 activity. The enhancement of SERCA2 activity triggered Ca2+ER perturbation, leading to an increase in autophagy. Thapsigargin blocked the interaction between SERCA2 and ER IGF-1Rβ and therefore SERCA2 activity, resulting in inhibition of HCC growth. In conclusion, the translocation of IGF-1R into the ER triggers Ca2+ER perturbation by enhancing SERCA2 activity through phosphorylating Tyr990 in HCC.

Activation of insulin-like growth factor-1 receptor (IGF-1R) promotes growth of colorectal cancer through triggering the MEX3A-mediated degradation of RIG-I

Insulin-like growth factor-1 receptor (IGF-1R) has been made an attractive anticancer target due to its overexpression in cancers. However, targeting it has often produced the disappointing results as the role played by cross talk with numerous downstream signalings. Here, we report a disobliging IGF-1R signaling which promotes growth of cancer through triggering the E3 ubiquitin ligase MEX3A-mediated degradation of RIG-I. The active β-arrestin-2 scaffolds this disobliging signaling to talk with MEX3A. In response to ligands, IGF-1Rβ activated the basal βarr2 into its active state by phosphorylating the interdomain domain on Tyr64 and Tyr250, opening the middle loop (Leu130‒Cys141) to the RING domain of MEX3A through the conformational changes of βarr2. The models of βarr2/IGF-1Rβ and βarr2/MEX3A could interpret the mechanism of the activated-IGF-1R in triggering degradation of RIG-I. The assay of the mutants βarr2Y64A and βarr2Y250A further confirmed the role of these two Tyr residues of the interlobe in mediating the talk between IGF-1Rβ and the RING domain of MEX3A. The truncated-βarr2 and the peptide ATQAIRIF, which mimicked the RING domain of MEX3A could prevent the formation of βarr2/IGF-1Rβ and βarr2/MEX3A complexes, thus blocking the IGF-1R-triggered RIG-I degradation. Degradation of RIG-I resulted in the suppression of the IFN-I-associated immune cells in the TME due to the blockade of the RIG-I-MAVS-IFN-I pathway. Poly(I:C) could reverse anti-PD-L1 insensitivity by recovery of RIG-I. In summary, we revealed a disobliging IGF-1R signaling by which IGF-1Rβ promoted cancer growth through triggering the MEX3A-mediated degradation of RIG-I.

LINC00649 Regulates Endoplasmic Reticulum Stress-mediated Cervical Carcinoma Cell Apoptosis via miR-424-5p/IGF1R Axis / 肿瘤防治研究

Objective To explore the effects of LINC00649/miR-424-5p/IGF1R on ERs-mediated apoptosis of cervical carcinoma (CC) cells. Methods CC-related data was obtained from GEO database, then the differentially-expressed miRNAs were analyzed. The bioinformatics database was used to predict the upstream and downstream targets of miR-424-5p. LINC00649 and IGF1R were included. Dual luciferase reporter assay was adopted to confirm the targeting relationship. qRT-PCR was used to detect the expression levels of LINC00649, miR-424-5p and IGF1R in CC tissue and cells. CCK-8 and flow cytometry were used to evaluate the proliferation and apoptosis of CC cells. Western blot was used to detect the expression of ERs-related proteins GRP78, CHOP and Caspase-12. Results Compared with paracancerous tissue and H8 cells, LINC00649 and IGF1R were up-regulated in CC tissue and cells, while miR-424-5p was down-regulated (both P < 0.05). The abnormally high expression of LINC00649 in CC was related to poor prognosis. The knockdown of LINC00649 inhibited CC cell viability and induced cell apoptosis by promoting ERs (all P < 0.05). LINC00649 upregulated the expression of IGF1R via absorbing miR-424-5p. miR-424-5p inhibitor or IGF1R overexpression partially reversed the effects of LINC00649 knockdown on CC cells (both P < 0.05). Conclusion LINC00649 could reduce cell apoptosis and improve cell viability by inhibiting the ERs process via regulating miR-424-5p/IGF1R axis in CC.

Porcine Circular RNA Insulin-like Growth Factor 1 Receptor Promotes Adipocyte Differentiation / 中国生物化学与分子生物学报

Circular RNA (circRNA), as a competitive endogenous RNA (ceRNA), plays an importantrole in the regulation of cell differentiation. The purpose of this study was to identify and analyze porcinecircular RNA insulin-like growth factor 1 receptor (circIGF1R), explore its expression patterns, construct a ceRNA regulatory network related to circIGF1R, and explore the regulation of its ectopicexpression on adipogenic differentiation of mouse mesenchymal stem cells (C3H10T1 / 2) effect. Forwardand reverse PCR, Sanger sequencing, RNase R enzyme digestion tests, and qRT-PCR were used toverify that circIGF1R is a circRNA formed by the second exon of insulin-like growth factor 1 receptor(IGF1R). It was expressed in all tissues of pigs, and its expression level increased with age in adiposetissues. miRDB, TargetScan and miRWalk online software were used to predict circIGF1R target genes. RNAhybrid software was used for binding site prediction. DAVID bioinformatics functional analysissoftware was used to perform GO and KEGG enrichment analysis on candidate target genes. Cytoscapesoftware was used to construct the ceRNA network diagram. Based on the gene expression correlation andpredicted target relationship, the GO and KEGG enrichment analysis was drawn and the ceRNA networkwas constructed; the dual luciferase reporter gene test was used, and we found that circIGF1R andFABP4 can bind to ssc (Sus scrofa chromosome) -miR-133a-5p. The circIGF1R overexpression vectorwas successfully constructed and expressed in C3H10T1/ 2 cells. It was found that after overexpression ofcircIGF1R, the expression of key adipogenic regulatory factors CEBPa, CEBPß, FABP4 and PPAR? increased significantly(P<0. 01), and the number of lipid droplets increased significantly. The results ofthis study show that circIGF1R exists in pig adipose tissues, and may positively regulate the adipogenicdifferentiation of C3H10T1/ 2 cells through the ceRNA mechanism, which lays a theoretical foundation forfurther research on circIGF1R regulating the adipogenic differentiation of pig precursor intramuscularadipocytes.

Over-expression of miR-145-5p inhibits malignant biological behaviors of esophageal squamous cells carcinoma via down-regulating IGF1R / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the mechanism of miR-145-5p on malignant biological behaviors, such as pro-liferation, invasion, migration and epithelial-mesenchymal transition (EMT), of esophageal squamous cell carcinoma (ESCC) TE-10 cells. Methods: The expression of miR-145-5p in ESCC cell lines and normal cells was detected by PCR. Dual luciferase reporter gene assay was used to detect the targeted regulation between miR-145-5p and insulin-like growth factor 1 receptor (IGF1R). The expres-sions of IGF1R protein and EMT related proteins were detected by Western blotting. Transwell assay and CCK-8 assay were carried out to detect the effects of miR-145-5p/IGF1R axis on the proliferation, migration andinvasionofTE-10 cells. Results: miR-145-5p was down-regulated in ESCC cell lines with the lowest expression in TE-10 cells (P<0.01orP<0.05).Over-expression of miR-145-5p significantly inhibited proliferation, invasion, migration and EMT of TE-10 cells (P<0.01 or P<0.05). Dual luciferase reporter gene assay con-firmed that miR-145-5p targetedly down-regulated IGF1R expression (P<0.01). The restora-tion experiments further confirmed that simultaneous over-expression of miR-145-5p and IGF1R significantly attenuated the promotion effect of IGF1R on proliferation, invasion, migration and EMT of TE-10 cells (P<0.01 or P<0.05). Conclusions: Over-expression of miR-145-5p inhibits proliferation, invasion, migration and EMT of ESCC TE-10 cells by down-regulating IGF1R.

Effect of interfering IGF-1R by siRNA on cell cycle and apoptosis of hypoxic hepatocellular carcinoma HepG2 cells / 中国肿瘤生物治疗杂志

@# Objective: To explore the effect of interfering insulin-like growth factors-1 receptors (IGF-1R) by small interfering RNA (siRNA) on cell cycle and apoptosis of hypoxic hepatocellular carcinoma HepG2 cells. Methods: The hypoxic hepatocellular carcinoma model was established via cobalt chloride treatment. Three siRNAs targeting IGF1R gene and one negative control siRNA were designed and synthesized. They were transfected into hypoxic HepG2 cells, and 24 h later, the transfection efficiency was detected by fluorescent microscopy. The protein expression of IFG-1R was detected with Western blotting (WB) to screen the siRNA with highest transfection efficacy. The selected siRNA was used to transfect hypoxic HepG2 cells. The proliferation of hypoxic HepG2 cells was determined by MTT assay. Cell cycle distribution and apoptosis were analyzed by Flow cytometry. WB was performed to detect the proteinexpressionsofCDK1,CDK2andCaspase-3inHepG2cells. Results: The hypoxic hepatocellular carcinoma model was successfully established. IGF-1R-siRNA-2 showed the most effective interference efficiency and the most significant knockdown of IGF-1R (all P<0.01). The proliferation of HepG2 cells transfected with IGF-1R siRNA-2 was significantly suppressed (P<0.05 or P<0.01), the cell cycle was blocked at G0/G1 phase (P<0.05), and the apoptosis rate was increased up to (25.3±1.3)% P<0.01). In the meanwhile, the expressions of CDK1 and CDK2 were decreased and the expression of Caspase-3 was increased in hypoxic HepG2 cells after IGF-1R knockdown (P<0.05). Conclusion: Interfering IGF-1R by siRNA inhibits the malignant biological behaviors of hypoxic HepG2 cells via regulating cell cycle and apoptosis-related proteins. IGF-1R may be a potential target for the treatment of HCC.

Expression and cellular distribution of insulin-like growth factor 1 receptor during window of implantation in infertile women with intramural fibroids

Background: The “window of implantation” (WOI) is a transient but well defined period during which the hostile endometrial lining is transformed to a surface receptive to accept the embryo. Recently, data are beginning to accumulate suggesting negative influence of non-cavity distorting intramural uterine fibroids (NCD-IMF) on endometrial receptivity that may have implications for implantation failure. However, molecular mechanisms underlying infertility associated with NCD-IMF remain unclear. The aim of present study was to examine the expression and cellular distribution of insulin-like growth factor 1 receptor (IGF1R) during WOI in infertile women with NCD-IMF and fertile controls. While, reports are available that support role of IGF1R in mediating adhesive interaction with the implanting blastocyst, the effect of NCD-IMF on IGF1R expression during the WOI is not defined.Methods: Quantitative real-time polymerase chain reaction and immunohistochemistry were used to evaluate messenger RNA (mRNA) and protein expression of IGF1R in midsecretory endometrial biopsies obtained from infertile women with NCD-IMF (n=20) and healthy fertile controls (n=10).Results: As compared to fertile controls, significantly higher IGF1R: i) mRNA levels (1.59 fold up regulation; p=0.044) and ii) immunoscore in the luminal epithelium (8.94±3.13 versus 6.31±1.49; p=0.009) were observed in infertile women with NCD-IMF.Conclusions: Over expression of IGF1R in infertile women with NCD-IMF, during the window of receptivity, may result in altered ability of uterine epithelial cells for blastocyst adhesion and subsequent implantation, which might lead to poor reproductive outcome in these women.

Novel mutation of type-1 insulin-like growth factor receptor (IGF-1R) gene in a severe short stature pedigree identified by targeted next-generation sequencing

Insulin-like growth factor receptor (IGF-1R) deficiency is a rare form of short stature, and is difficult to clinically diagnose. Targeted next-generation sequencing (NGS) allows for the rapid and inexpensive assessment of short stature. We identified mutations in the pedigree of a Chinese boy with severe short stature using targeted NGS; we then assessed the clinical characteristicsand evaluated the efficacy of growth hormone therapy. NGS analysis revealed a novel heterozygous missense mutation in exon3 (c.926C>T, p.S309L) of the type-I IGF-1R gene in the proband, which was inherited from the mother. The proband, mother and grandfather suffered from severe growth failure. After recombinant human growth hormone therapy, the patient’s growth rate increased. The novel missensemutation in IGF-1R (c.926C > T, p.S309L) is associated with severe short stature in Chinese individuals. Targeted NGS may enable efficient diagnosis and genetic consultation of children with short stature.

IGF-1R/β-catenin signaling axis is involved in type 2 diabetic osteoporosis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Insulin-like growth factor-1 receptor (IGF-1R) is involved in both glucose and bone metabolism. IGF-1R signaling regulates the canonical Wnt/β-catenin signaling pathway. In this study, we investigated whether the IGF-1R/ β-catenin signaling axis plays a role in the pathogenesis of diabetic osteoporosis (DOP). Serum from patients with or without DOP was collected to measure the IGF-1R level using enzyme-linked immunosorbent assay (ELISA). Rats were given streptozotocin following a four-week high-fat diet induction (DOP group), or received vehicle after the same period of a normal diet (control group). Dual energy X-ray absorption, a biomechanics test, and hematoxylin-eosin (HE) staining were performed to evaluate bone mass, bone strength, and histomorphology, respectively, in vertebrae. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were performed to measure the total and phosphorylation levels of IGF-1R, glycogen synthase kinase-3β (GSK-3β), and β-catenin. The serum IGF-1R level was much higher in patients with DOP than in controls. DOP rats exhibited strikingly reduced bone mass and attenuated compression strength of the vertebrae compared with the control group. HE staining showed that the histomorphology of DOP vertebrae was seriously impaired, which manifested as decreased and thinned trabeculae and increased lipid droplets within trabeculae. PCR analysis demonstrated that IGF-1R mRNA expression was significantly up-regulated, and western blotting detection showed that phosphorylation levels of IGF-1R, GSK-3β, and β-catenin were enhanced in DOP rat vertebrae. Our results suggest that the IGF-1R/β-catenin signaling axis plays a role in the pathogenesis of DOP. This may contribute to development of the underlying therapeutic target for DOP.

IGF-1R/β-catenin signaling axis is involved in type 2 diabetic osteoporosis / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Insulin-like growth factor-1 receptor (IGF-1R) is involved in both glucose and bone metabolism. IGF-1R signaling regulates the canonical Wnt/β-catenin signaling pathway. In this study, we investigated whether the IGF-1R/ β-catenin signaling axis plays a role in the pathogenesis of diabetic osteoporosis (DOP). Serum from patients with or without DOP was collected to measure the IGF-1R level using enzyme-linked immunosorbent assay (ELISA). Rats were given streptozotocin following a four-week high-fat diet induction (DOP group), or received vehicle after the same period of a normal diet (control group). Dual energy X-ray absorption, a biomechanics test, and hematoxylin-eosin (HE) staining were performed to evaluate bone mass, bone strength, and histomorphology, respectively, in vertebrae. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were performed to measure the total and phosphorylation levels of IGF-1R, glycogen synthase kinase-3β (GSK-3β), and β-catenin. The serum IGF-1R level was much higher in patients with DOP than in controls. DOP rats exhibited strikingly reduced bone mass and attenuated compression strength of the vertebrae compared with the control group. HE staining showed that the histo-morphology of DOP vertebrae was seriously impaired, which manifested as decreased and thinned trabeculae and increased lipid droplets within trabeculae. PCR analysis demonstrated that IGF-1R mRNA expression was significantly up-regulated, and western blotting detection showed that phosphorylation levels of IGF-1R, GSK-3β, and β-catenin were enhanced in DOP rat vertebrae. Our results suggest that the IGF-1R/β-catenin signaling axis plays a role in the pathogenesis of DOP. This may contribute to development of the underlying therapeutic target for DOP.

Silencing SPARC inhibits high fluoride-induced apoptosis of thyrocytes / 西安交通大学学报(医学版)

Objective: To investigate the effects and mechanisms of secreted protein, acidic and rich in cysteine (SPARC) on high fluoride-induced apoptosis of thyrocytes. Methods: Human thyroid cells (Nthy-ori 3-1) were cultured and treated with various concentrations (0.1, 1 and 10 mmol/L) of NaF for 24 h, and the expression of SPARC was evaluated using Real-time PCR and Western blot, respectively. The cells were divided into four groups: control group, NaF group, si-SPARC group (cells were transfected with SPARC siRNA for 48 h and then exposed to NaF for 24 h), and si-NC group (cells were transfected with negative control siRNA for 48 h and then exposed to NaF for 24 h). Cytotoxicity was assayed using CCK-8 and LDH; cell apoptosis rate was detected with ELISA. The expressions of cleaved caspase3 (c-caspase3) and IGF-1R were measured by Western blot. In addition, si-SPARC and si-IGF-1R were co-transfected into thyrocytes to further explore mechanisms of SAPRC by evaluating apoptosis. Results: The mRNA and protein levels of SPARC were augmented with the increase of NaF (P<0.05). Cell viability was significantly higher in si-NC group than that in si-SPARC group [(84.02±9.51)% vs. (58.31±6.86)%, P<0.05], and the release rate of LDH was lower [(134.25±18.98)% vs. (195.18±23.50)%, P<0.05]. Cell apoptosis rate was lower in si-SPARC group than that in si-NC group [(124.67±19.44)% vs. (175.24±16.46)%, P<0.05]. In addition, silencing SPARC upregulated the expression of IGF-1R (1.95±0.24 vs. 0.93±0.08, P<0.05), and inhibition of IGF-1R reversed the effect of SPARC on apoptosis. Conclusion: Inhibition of SPARC reduces high fluoride-induced cytotoxicity and blocks cell apoptosis. The possible mechanism is through the negative regulation of IGF-1R.

Downregulation of lncRNA CCDC26 contributes to imatinib resistance in human gastrointestinal stromal tumors through IGF-1R upregulation

Imatinib is the first line of therapy for patients with metastatic or gastrointestinal stromal tumors (GIST). However, drug resistance limits the long-term effect of imatinib. Long non-coding RNAs (lncRNAs) are emerging as key players in regulating drug resistance in cancer. In this study, we investigated the association between lncRNA CCDC26 and IGF-1R in GIST and their involvement in drug resistance. Considering the key role of lncRNAs in drug resistance in cancer, we hypothesized that IGF-1R is regulated by lncRNAs. The expression of a series of reported drug resistance-related lncRNAs, including CCDC26, ARF, H19, NBR2, NEAT1, and HOTAIR, in GIST cells treated with imatinib H19 was examined at various time-points by qRT-PCR. Based on our results and published literature, CCDC26, a strongly down-regulated lncRNA following imatinib treatment, was chosen as our research target. GIST cells with high expression of CCDC26 were sensitive to imatinib treatment while knockdown of CCDC26 significantly increased the resistance to imatinib. Furthermore, we found that CCDC26 interacted with c-KIT by RNA pull down, and that CCDC26 knockdown up-regulated the expression of IGF-1R. Moreover, IGF-1R inhibition reversed CCDC26 knockdown-mediated imatinib resistance in GIST. These results indicated that treatments targeting CCDC26-IGF-1R axis would be useful in increasing sensitivity to imatinib in GIST.

Effects of electroacupuncture at "Zhongliao" (BL 33) and "Tianshu" (ST 25) on ovarian function in rats with premature ovarian insufficiency / 中国针灸

<p><b>OBJECTIVE</b>To explore the effects of electroacupuncture (EA) at "Zhongliao" (BL 33) and "Tianshu" (ST 25) on ovarian function in rats with premature ovarian insufficiency (POI).</p><p><b>METHODS</b>A total of 48 SD female rats with regular estrus were divided into a blank group (=8), a model group (=10), an EA group (=10), a binding group (=10) and a tamoxifen (TAM) group (=10). The rats in the model group, EA group, binding group and TAM group were all treated with intraperitoneal injection of 4-vinylcyclohexene diepoxide (VCD, 160 mg/kg) for 15 consecutive days to establish the model of POI; the rats in the blank group were treated with normal diet. After the model was established successfully, the rats in the EA group were treated with EA at "Zhongliao" (BL 33) and "Tianshu" (ST 25) with continuous wave (1 to 3 Hz, 0.1 to 1 mA) for 20 minutes, once a day (five times a week) for the first two weeks and once every other day (three times a week) for the following two weeks. The rats in the TAM group were treated with subcutaneous injection of tamoxifen (1mg/kg), once a day (five times a week) for the first two weeks and once every other day (three times a week) for the following two weeks. The rats in the binding group were bound by a small sack as the EA group. The rats in the blank group and the model group were treated with normal diet. After four weeks, the sexual gland weight and index were tested in each group; the ELISA method was applied to test the level of anti-mllerian hormone (AMH) and inhibin B; the morphology of ovary was observed; the number of primordial follicles, primary follicle, antral follicle and atretic follicle was counted; the expression of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-1 receptor (IGF-1R) were measured.</p><p><b>RESULTS</b>(1) Compared with the blank group, the ovary weight, ovary index, uterus weight and uterus index were significantly decreased after treatment in the model group, EA group, binding group and TAM group (all <0.01); but the differences between the model group and the EA group, binding group, TAM group were not significant (all >0.05). (2) Compared with the blank group, the levels of serum AMH, inhibin B and E were significantly reduced; the levels of FSH and LH were significantly increased in the model group; EA group, binding group and TAM group (all <0.01). Compared with the model group, the levels of serum AMH, inhibin B and E were significantly increased, the level of FSH and LH were significantly reduced in the EA group and TAM group (all <0.01). (3) Compared with the blank group, in the model group, EA group, binding group and TAM group the ovary was dark red and pale, surrounded by particle or not; the morphology was small and atrophic; the primordial follicles was reduced even vanished; the structure of primary follicle was damaged and loosely arranged; the mature follicle was few; the atretic follicle and interstitial gland were increased. (4) Compared with the blank group, the expressions of IGF-1 mRNA and IGF-1R mRNA were increased in the model group (all <0.01); compared with the blank group, the expression of IGF-1 mRNA was increased in the binding group (<0.05), but that of IGF-1R mRNA was not significantly different (>0.05); compared with the model group, the expression of IGF-1 mRNA was not significantly different in the EA group, binding group and TAM group (all >0.05), but that of IGF-1R mRNA was reduced (<0.05, <0.01).</p><p><b>CONCLUSION</b>EA at "Zhongliao" (BL 33) and "Tianshu" (ST 25) has improvement effect on ovarian function in rats with VCD-induced POI, which is likely to be related to regulating the IGF-1R mRNA expression to improve the IGF-1/ IGF-1R axis.</p>

Expression and Clinical Significance of IGF-1R and VEGFR2 Protein in Colorectal Cancer Tissue / 中国药房

OBJECTIVE:To observe the expression of IGF-1R and VEGFR2 in colorectal cancer tissue,and to explore its clinical significance. METHODS:Totally 98 colorectal cancer patients underwent surgery were selected from our hospital during Jan. 2011-Dec. 2012. The tumor tissue and paracancerous tissue were collected. The expression of IGF-1R and VEGFR2 protein were detect-ed by immunohistochemical SP method. The mRNA expression of both were detected by RT-PCR. The correlation of the expression of IGF-1R and VEGFR2 protein with clinical pathological characteristics of colorectal cancer patients were investigated,and its effects on 4-year disease-free survival rate of postoperative assisted chemotherapy patients were also investigated. RESULTS:The positive rates of IGF-1R and VEGFR2 protein expression in colorectal cancer tissue were 66.3％and 60.2％respectively,but their expression was not found in paracancerous tissue,with statistical significance(P<0.05). Relative expression of IGF-1R mRNA and VEGFR2 mRNA in tumor tissue were significantly higher than paracancerous tissue,with statistical significance(P<0.05). There was positive correla-tion between their mRNA expression in tumor tissue(r=0.729,P<0.05). The expression of IGR-1R protein was independent from pa-tient's gender and age,tumor location and formation of vascular tumor thrombus(P>0.05);but it was associated with the histological grading,depth of invasion and lymph node metastasis(P<0.05). The expression of VEGFR2 protein had nothing to do with patient's gender and age(P>0.05),but it was associated with tumor location,histological grading,depth of invasion,lymph node metastasis, and formation of vascular tumor thrombus(P<0.05). Among 98 colorectal cancer patients,14 patients didn't underwent chemothera-py after surgery;and 4-year disease-free survival rate of other 84 patients was 76.2％(64/84). Four-year disease-free survival rates of TGF-1R protein negative expression and VEGFR2 protein negative expression patients(82.8％,78.8％)were significantly higher than those of positive expression patients(72.7％,74.5％),with statistical significance(P<0.05). The multiple factor Logistic regression analysis was used to correct gender,age,histological grading,tumor location,depth of invasion,lymph node metastasis,and forma-tion of vascular tumor thrombus;the expression of IGR-1R and VEGFR2 protein were still associated with 4-year disease-free survival rate [adjusted OR were 2.31,2.15;95％CI were(0.57,4.15),(0.45,4.03);P<0.05]. CONCLUSIONS:The expression of IGF-1R, VEGFR2 protein and mRNAs increase significantly in colorectal cancer tissue;the protein expression is associated with histological grading,depth of invasion,lymph node metastasis and the prognosis of postoperative assisted chemotherapy patients. Their examina-tion can provide reference for target therapy for colorectal cancer patients.

Implications of Insulin-like Growth Factor 1 Receptor Activation in Lung Cancer

Insulin-like growth factor 1 receptor (IGF1R) has been intensively investigated in many preclinical studies using cell lines and animal models, and the results have provided important knowledge to help improve the understanding of cancer biology. IGF1R is highly expressed in patients with lung cancer, and high levels of circulating insulin-like growth factor 1 (IGF1), the main ligand for IGF1R, increases the risk of developing lung malignancy in the future. Several phase I clinical trials have supported the potential use of an IGF1R-targeted strategy for cancer, including lung cancer. However, the negative results from phase III studies need further attention, especially in selecting patients with specific molecular signatures, who will gain benefits from IGF1R inhibitors with minimal side effects. This review will discuss the basic concept of IGF1R in lung cancer biology, such as epithelial-mesenchymal transition (EMT) induction and cancer stem cell (CSC) maintenance, and also the clinical implications of IGF1R for lung cancer patients, such as prognostic value and cancer therapy resistance.

SilencingIGF-1R gene inhibits proliferation of human SMMC7721 cell and promotes its apoptosis through down-regulatingBMP2 expression / 中国癌症杂志

Background and purpose:Insulin-like growth factor-1 (IGF-1) is a peptide that participates in many biological processes by stimulating the downstream signaling pathways through their interaction with IGF-1 re-ceptor (IGF-1R) and insulin receptor (IR). Bone morphogenetic proteins (BMPs) are a group of functional proteins which participate in the biological processes of proliferation and migration in many kinds of cancers and have become a hot area of cancer research. The study aimed to investigate the effects of silencingIGF-1R gene on the expression level ofBMP2 gene, and the cell proliferation and apoptosis of SMMC7721 cells.Methods:The RNAi plasmid targetingIGF-1R gene was constructed and transfected into SMMC7721 cells. Then the inhibition effect on the expression level of IGF-1R and BMP2 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The SMMC7721 growth curve and cell apoptosis were detected by MTT assay and flow cytometry after they were transfected with RNAi plasmid.Results:The RNAi plasmid targetingIGF-1R gene was constructed successfully. The inhibition efficiencies at mRNA expression levels were 68.9% and 80.7% (IGF-1R gene), 79.5% and 83.3% (BMP2 gene), respectively, after transfection with IGF-1R-siRNA-1 and IGF-1R-siRNA-2 plasmid (P<0.05). The inhibition efficiencies at protein levels were 46.1% and 62.1% (IGF-1R gene,P<0.05), 42.5% and 60.9% (BMP2 gene,P<0.05), respectively. The results of MTT growth curve showed that the proliferation rate in the transfected SMMC7721 cells was significantly slower than that in the control group (P<0.05). The proportion of apoptotic cells in transfected groups was significantly higher than that in the control group (P<0.05).Conclusion:SilencingIGF-1R gene can downregulate the expression ofBMP2 gene at different levels that results in inhibition of cell proliferation and promotion of apoptosis in SMMC7721 cells.

Bioinformatics analysis of sequence and structure of insulin-like growth factor-I receptor / 生物工程学报

The length of IGF1R 3'UTR is greater than 7 kb. The structure of IGF1R 3'UTR is complex, with multiple binding sites of miRNAs. IGF1R is involved in the regulation of MAPK and PI3K/AKT signaling pathways and theformation and development of tumors. Bioinformatics analysis can reveal the structure features of IGF1R, which provides ideas for further research. The analysis shows that the binding sites between IGF1R and miRNAs have the highest mutation rate in Neuroblastoma. We analyzed the structure of 3'UTR, miRNAs binding sites, physical and chemical properties, hydrophilic-hydrophobic property, glycosylation and phosphorylation sites, secondary structure and tertiary structure modeling of IGF1R. The locations and names of amino acids interacting in IGF1R and IGF1 were obtained by molecular docking. Therefore, if IGF1R 3'UTR is mutated, the capacity of IGF1R combined with miRNAs will reduce and the IGF1R expression will be up-regulated, and the function of miRNAs will be repressed. We can change the sites of IGF1R to combine with IGF1 to repress the function of IGF1R and IGF1. Then the function of IGF1R will be repressed.

The relationship between expression of E -cad and IGF -1R and the recurrence of patients with HCC after liver resection / 中国基层医药

Objective To investigate the relationship between expression of E -cad and IGF -1R and the recurrence of patients with primary hepatocellular carcinoma (HCC)who underwent liver resection.Methods The clinical data of 91 cases of primary HCC underwent liver resection from January 2010 to December 2012 were analyzed retrospectively.The expression of E -cad and IGF -1R was detected by RT -PCR,and the relationship between expression of E -cad and IGF -1R and the recurrence of patients was analyzed.Results The expression of E -cad was obviously lower in HCC tissues than that in normal liver tissues,with the expression percentage of 38.7% vs. 75.0%.The recurrence rate of E -cad negative group was higher(71.1%).The TNM Ⅱ -Ⅳ stage (χ2 =7.161, P =0.009)and liver capsule invasion (χ2 =5.144,P =0.036)in E -cad negative group were higher than E -cad positive group.The expression of IGF -1R was obviously higher in HCC tissues than that in normal liver tissues,with the expression percentage of 66.1% vs.20.0%.The recurrence rate of IGF -1R negative group was higher(68.3%). The TNMⅡ-Ⅳ stage (χ2 =4.195,P =0.014),liver capsule invasion (χ2 =5.144,P =0.036),non -tumor capsularin (χ2 =7.201,P =0.012)and PVTT(χ2 =6.538,P =0.032)in E -cad negative group were higher than E -cad positive group.And there was negative correlation between the expression of E -cad and IGF -1R(χ2 =14.329,P =0.000).Conclusion The mRNA expression of E -cad in the tissues of HCC is obviously lower than normal liver tissues.The mRNA expression of IGF -1R in the tissues of HCC is obviously higher than normal liver tissues.The expressions of E -cad and IGF -1R are associated with the degree of local invasion and malignant.The E -cad and IGF -1R expression is correlated in the tissues of HCC,which means that E -cad and IGF -1R play synergistic effect in HCC genesis.

Expressions and clinical significances of IGF-1R and IRS-1 in squamous cell lung cancer / 实用肿瘤学杂志

Objective The aim of the study is to detect the expression of IGF -1R and IRS-1 in squa-mous cell lung cancer ( SQCLC) and to explore the role in the development of squamous cell lung cancer and clini -cal significance .Methods Specimens from 246 surgical SQCLC and 40 adjacent normal lung tissues were evalu-ated for IGF-1R and IRS-1 expression by immunohistochemistry .We explored the correlation between IGF -1R and IRS-1,their relationship with clinicopathological parameters and their impact on outcome in SQCLC pa -tients.Results SQCLC of IGF -1R positive expression rate was 54.07%,which was higher than in adjacent normal tissue(32.5%),SQCLC of IRS-1 positive expression rate was 38.21%,which was lower than in adja-cent normal tissue(70%),the pairwise difference was statistically significant (P<0.05);Expression of IGF-1R was correlated with lymph node metastasis (P<0.05),IRS-1 expression was related with degree of differentia-tion and lymph node metastasis (P<0.05).Survival of IGF -1R expression group of patients was significantly shorter than the expression of IGF -1R negative patients ,the survival of the IRS-1 expression was significantly longer than patients who negatively expressed IRS -1,the difference was statistically significant (P=<0.001 and=0.021).IGF-1R was negatively correlated with IRS -1 expression in SQCLC(r=-0.125,P<0.001). Conclusion IGF-1R,IRS-1 are involved in the occurrence of SQCLC ,the development .The IGF-1R is an independent prognostic factor in SQCLC .