P4HA2 promotes occurrence and progression of liver cancer by regulating the PI3K/Akt/mTOR signaling pathway / 南方医科大学学报

OBJECTIVE@#To investigate the role of proline 4-hydroxylase Ⅱ (P4HA2) in the occurrence and progression of liver cancer.@*METHODS@#GEPIA and Human Protein Atlas database were used to predict the expression of P4HA2 in hepatocellular carcinoma (HCC), and K-M plotter online database was used to analyze the relationship between P4HA2 expression and the prognosis of HCC. We also examined the expressions of P4HA2 in HCC cells and normal hepatocytes using qRT-PCR and Western blotting. With lentivirus-mediated RNA interference, P4HA2 expression was knocked down in hepatoma SNU-449 and Hep-3B cells, and the changes in cell proliferation, migration and invasion were assessed using cell counting kit-8 (CCK-8) assay, colony formation test, scratch test and Transwell assay. The changes in the expressions of epithelial-mesenchymal transition (EMT) and PI3K/Akt/mTOR signal pathway-related proteins were detected using Western blotting.@*RESULTS@#Online database analysis showed that the expression of P4HA2 was significantly higher in HCC tissues than in normal liver tissues (P < 0.05). The expression levels of P4HA2 mRNA and protein were also significantly higher in HCC cell lines than in normal hepatocytes (P < 0.01). Lentivirus-mediated RNA interference of P4HA2 significantly lowered the expression levels of P4HA2 mRNA and protein in the hepatoma cells (P < 0.05) and caused obvious inhibition of cell proliferation, migration and invasion. P4HA2 knockdown significantly increased the expression of E-cadherin protein, lowered the expressions of N-cadherin and Snail, and obviously decreased the expressions of phosphorylated PI3K, AKT and mTOR (P < 0.05).@*CONCLUSION@#P4HA2 enhances the proliferation, migration, invasion, and EMT of hepatoma cells by activating the PI3K/Akt/mTOR signaling pathway to promote the occurrence and progression of liver cancer.

Inhibition effect of active fraction from clove on PI3K/Akt/mTOR signaling pathway to induce apoptosis of human colon cancer HCT116 cells / 中国中药杂志

To screen the sensitive cell lines of active fraction from clove(AFC) on human colon cancer cells, investigate the effects of AFC on the cells proliferation and apoptosis as well as PI3 K/Akt/mTOR(phosphoinositide 3-kinase/Akt/mechanistic target of rapamycin) signaling pathways involved, and reveal the mechanism of AFC for inducing apoptosis of human colorectal carcinoma cells. Cell counting kit-8(CCK-8) assay was used to detect the cytotoxic effect of different concentrations of AFC. AFC-induced apoptosis was detected by Hoechst 33258 fluorescence staining and Annexin V-FITC/PI double staining. HCT116 cells were treated with AFC with or without pretreatment with insulin-like growth factor-Ⅰ(IGF-Ⅰ), and then the protein expression levels of caspase-3, caspase-9, poly ADP-ribose polymerase(PARP), PI3 K, p-PI3 K, Akt, p-Akt, mTOR and p-mTOR in PI3 K/Akt/mTOR signaling pathway were detected by Western blot. RESULTS:: showed that the most obvious inhibitory effect of AFC was on human colon cancer HCT116 cells, and the optimal AFC treatment time was 48 hours. After AFC treatment, typical apoptotic features such as nuclear chromatin concentration, nuclear fragmentation and apoptotic bodies appeared in a dose-dependent manner. Annexin V-FITC/PI double staining showed that as compared with the control group, 50 and 100 μg·mL~(-1) AFC groups increased the apoptosis rate of HCT116 cells significantly(P<0.001); AFC activated caspase-9, cleaved caspase-3 and cleaved PARP in a concentration-dependent manner. The protein expression levels of cleaved caspase-3/procaspase-3, cleaved PARP/PARP and caspase-9/β-actin after treatment of AFC(100 μg·mL~(-1)) were significantly different from those in the control group(P<0.001). The relative protein expression of p-PI3 K, p-Akt and p-mTOR decreased in a concentration dependent manner, while Akt and mTOR showed no significant differences among groups. The ratios of p-PI3 K/PI3 K, p-Akt/Akt and p-mTOR/mTOR in the AFC groups(50 and 100 μg·mL~(-1)) were significantly lower than those in the control group(P<0.01). Its combination with IGF-Ⅰ weakened the effect of AFC in inhibiting PI3 K/Akt/mTOR signaling pathway. The ratios of p-Akt/Akt and p-mTOR/mTOR in the AFC+IGF-Ⅰ group were significantly enhanced as compared with the AFC group(P<0.05). Apoptosis-related protein expression levels(cleaved caspase-3 and cleaved PARP) in HCT116 cells treated with AFC+IGF-Ⅰ were also down regulated. As compared with the AFC group, the ratios of cleaved caspase-3/procaspase-3 and cleaved PARP/PARP in the AFC+IGF-Ⅰ group were significantly decreased(P<0.01). In summary, AFC activated caspase-mediated cascades and induced HCT116 cells apoptosis in a dose-dependent manner, which may be associated with the inhibition of the PI3 K/Akt/mTOR signaling pathway.

Mulberry polysaccharides improves the chemotherapy of liver cancer ascites tumor-bearing mice by regulating the PI3K/AKT/mTOR pathway / 中国临床药理学与治疗学

AIM: To study the synergistic and attenuating effects of mulberry polysaccharides on the chemotherapy of liver cancer ascites tumor-bearing mice by regulating the PI3K/AKT/mTOR pathway. METHODS: Ninety SPF male Kunming mice were randomly divided into normal group, model group, cyclophosphamide group, mulberry polysaccharide group and mulberry polysaccharide + cyclophosphamide group. Liver cancer ascites tumor-bearing mice were prepared, and each administration group was given 30 mg/kg of cyclophosphamide or (and) 200 mg/kg of mulberry polysaccharide and administered orally. The normal group and the model group were administrated with 10 mL/kg saline. The tumor suppression rate, liver, spleen and other indexes, tumor tissue VEGF and inflammatory factor content, and PI3K/AKT/mTOR pathway-related protein expression were observed in mice.RESULTS:Cyclophosphamide group, mulberry polysaccharide group and mulberry polysaccharide + cyclophosphamide group mice body weight, tumor mass, tumor tissue VEGF, TNF-α, IL-6, PI3K, AKT and mTOR phosphorylation levels were lower than the model group, and the level of IL-1β was higher than the model group; mulberry polysaccharide + cyclophosphamide group mice were observed with body weight, tumor inhibition rate, tumor tissue VEGF, TNF-α, IL-6, PI3K, AKT and The mTOR phosphorylation level higher than the mulberry polysaccharide group and cyclophosphamide group, and the tumor mass and IL-1β level were lower than the mulberry polysaccharide group and cyclophosphamide group (P<0.05). CONCLUSION: Morus alba polysaccharide combined with cyclophosphamide can effectively inhibit tumor proliferation of liver cancer ascites tumor-bearing mice and exert attenuating effect. The mechanism may be related to the down-regulation of PI3K /AKT/mTOR pathway expression.

Polydatin protects H9c2 cells from hypoxia-induced injury via up-regulating long non-coding RNA DGCR5

Polydatin (PD), a monocrystalline polyphenolic drug mainly found in the roots of Polygonum cuspidatum, has various pharmacological activities. Long non-coding RNAs (lncRNA) DiGeorge syndrome critical region gene 5 (DGCR5) was found to participate in the suppression of multiple cancers. Here, we proposed to study the effect of PD on myocardial infarction (MI) by inducing DGCR5. CCK-8 assay was performed to detect the viability of H9c2 cells. Flow cytometry was utilized to test apoptosis of H9c2 cells. These results determined the optimal concentration and effect time of hypoxia as well as PD. Si-DGCR5 was transfected into cells and the expression level was determined by qRT-PCR. Western blot was utilized to evaluate the expression of apoptosis-related proteins, Bcl-2, Bax, and cleaved-caspase-3, as well as autophagy-associated proteins including Beclin-1, p62, and LC3-II/LC3-I. As a result, PD efficiently attenuated hypoxia-induced apoptosis and autophagy in H9c2 cells. The expression of DGCR5 was down-regulated by hypoxia and up-regulated by PD. Besides, knocking-down the expression of DGCR5 inhibited the protection of PD in H9c2 cells. In addition, PD up-regulated the accumulation of DGCR5, DGCR5 decreased the expression of Bcl-2 and p62, raised the expression of Bax and cleaved-caspase-3, and the proportion of LC3-II/LC3-I. PD stimulated the PI3K/AKT/mTOR and MEK/ERK signaling pathways via up-regulating the expression of DGCR5. Our data demonstrated that PD reduced cell apoptosis and autophagy induced by hypoxia in cardiomyocytes. Moreover, PD activated PI3K/AKT/mTOR and MEK/ERK signaling pathways by up-regulating the expression of DGCR5.

Inhibition of peiminine on invasion and migration of human lung cancer A549 cells by decreasing epithelial-mesenchymal transition process via PI3K/Akt/mTOR pathway / 中草药

Objective: To detect the influence of peiminine on the invasion and migration of human lung cancer A549 cells. Methods A549 cells were treated with peiminine at the final concentrations of 0, 50, 100, and 200 μmol/L, respectively. The influence of peiminine on the invasion and migration of A549 cells and its underlying mechanisms were investigated by cell invasion experiment, cell scratch experiment, real-time quantitative PCR, ELISA, and Western blotting. Results Compared with 0 μmol/L peiminine group, the cell transmembrane number and scratch wound healing rate and expressions of MMP-9 and MMP-2 in the group treated with different concentrations of peiminine significantly decreased (P < 0.01). Compared with 0 μmol/L peiminine group, the mRNA expression of E-cadherin increased significantly (P < 0.01), while the mRNA expressions of N-cadherin and vimentin decreased significantly (P < 0.01). Compared with 0 μmol/L peiminine group, FN protein expression was significantly decreased in all the groups with different concentrations of peiminine group except treatment with 50 μmol/L peiminine after 24 h (P < 0.05, 0.01). Compared with 0 μmol/L peiminine group, the ratio of p-PI3K/PI3K and p-mTOR/mTOR in all concentrations of peiminine groups and p-Akt/Akt in 100 and 200 peiminine groups were significantly decreased (P < 0.05, 0.01). Conclusion Peiminine can inhibit the invasion and migration of A549 cells, which may be related to the activation of PI3K/Akt/mTOR pathway and decreasing the epithelial- mesenchymal transition process.

Pharmacological analysis of curdione on hypertrophic scar fibroblast / 中草药

Objective To investigate the effects of curdione on the HSF proliferation, transformation, and collagen secretion. Methods After the human HSF was treated with curdione, the proliferation inhibition ratio was measured using MTT method. Meanwhile, the TIMP-1, MMP-1, COL-I, and COL-III were detected by ELISA method, the α-SMA was analyzed by IHC technology, and the PI3K/Akt/mTOR and TGF-β1/Smads related molecular were evaluated by Western blotting. Results Curdione could reduce the proliferation inhibition ratio. Compared with control group, the TIMP-1, COL-I, and COL-III secretion were inhibited by curdione significantly (P < 0.05, P < 0.01), while the MMP-1 levels was significantly increased by curdione (P < 0.05, P < 0.01). The results also indicated that the expression levels of p-PI3K, p-Akt, p-mTOR, p-Smad3, TGF-β1, and α-SMA were significantly suppressed by curdione with concentration dependence (P < 0.05, P < 0.01). Conclusion Curdione could effectively improve the hypertrophic scar by inhibiting the HSF proliferation, transformation, and collagen secretion, and accelerating the collagen enzymolysis via PI3K/Akt/mTOR and TGF-β1/Smads pathways.

Mechanism research of safflower polysaccharide inducing apoptosis of human breast cancer MDA-MB-435 cells by interrupting PI3K/Akt/mTOR Pathway / 中草药

Objective To investigate the effect of safflower polysaccharide on apoptosis of human breast cancer MDA-MB-435 cells by blocking PI3K/Akt/mTOR pathway and explore its mechanism. Methods MDA-MB-435 cells were divided into control group and safflower polysaccharide 0.5 and 1.0 mg/mL groups. MTT assay and flow cytometry were used to detect the effects of different concentrations of safflower polysaccharide on the growth inhibition and apoptosis of MDA-MB-435 cells. RT-PCR and Western blotting were used to detect the effect of different concentrations of safflower polysaccharides on the mRNA and protein expression of PI3K, Akt, and mTOR in MDA-MB-435 cells. Results Compared with the control group, the inhibition rate of MDA-MB-435 cells in safflower polysaccharide 1.0 mg/mL group was (27.73 ± 3.75)%, which was significantly higher than that [(21.52 ± 2.43)%] in safflower polysaccharide 0.5 mg/mL group (P < 0.05). Compared with the control group, safflower polysaccharide could significantly increase the apoptosis rate of MDA-MB-435 cells in a dose-dependent manner (P < 0.01). The results of RT-PCR and Western blotting showed that, compared with the control group, safflower polysaccharide significantly decreased the mRNA and protein expression of PI3K, Akt, and mTOR in MDA-MB-435 cells (P < 0.05, 0.01). Conclusion Safflower polysaccharide can effectively inhibit the growth of MDA-MB-435 cells and promote their apoptosis, which may be achieved by blocking the PI3K/Akt/mTOR pathway.

Effect and mechanism of Stellettin B induced human glioblastoma SF295 cells autophagy / 国际生物医学工程杂志

Objective To study the autophagy of human glioblastoma SF295 cells induced by sponge-derived triterpenoid Stellittin B (Stel B) and to discuss the related mechanism. Methods The proliferation inhibitory activity of Stel B on SF295 cells was studied by WST-8 assay. The autophagy effect induced by Stel B on SF295 cells was evaluated bya variety of experimental techniques, including MDC staining, Western Blot assay, and mRFP-GFP-LC3 plasmid transfection method.The activity of Stel B on the representative signal proteins in PI3K/Akt/mTOR pathway in SF295 cells was examined by Western Blot. The anti-tumor activity of Stel B was detected by WST-8 assay after blocking autophagy by autophagy inhibitor chloroquine. Results Stel B could significantly inhibit the proliferation of SF295 cells with IC50 value as 0.026 μmol/L. Plenty of autophagosome was found in Stel B treated A549 cells by MDC staining. The expression of autophagy marker proteins including LC3B-II increased. The confocal microscopy results showed that Stel B promoted autophagosome forming and inhibitedthe fusion of autophagosome with lysosome. The Western Blot results showed that Stel B inhibited the expression of PI3K-p110 protein in SF295 cells and decreased the phosphorylation level of Akt and mTOR.The inhibitory effects of Stel B with different concentrations on SF295 cells were all enhanced by the inhibition of autophagy with chloroquine, and the differences were statistically significant (all P<0.05). Conclusion Stel B can induce SF295 cells autophagy via blocking PI3K/Akt/mTOR pathway. This autophagy effect is beneficial to the survival of SF295 cells. The antitumor activity of Stel B can be enhanced by a combination of autophagy inhibitors.

Expression of PSAT1 in pancreatic cancer tissues and the mechanism underlying PSAT1-mediated cell proliferation and invasion / 中国肿瘤临床

Objectives: To investigate the expression of phosphoserine aminotransferase 1 (PSAT1) in pancreatic cancer tissues, and its potential role in pancreatic cancer. Methods: The expression of PSAT1 in 98 human pancreatic cancer tissues, which were collected from the People＇s Hospital of Guizhou, between July 2013 to July 2017, and the corresponding adjacent normal tissues was analyzed by immunohistochemical staining. Additionally, the relationship between the expression of PSAT1 and the clinicopathological parame-ters, overall survival (OS), and disease-free survival (DFS) of patients with pancreatic cancer was evaluated. The human pancreatic can-cer cell lines, BxPC-3 and SW1990, were transfected with PSAT1-siRNA, to investigate the effect of PSAT1 knockdown on cell prolifera-tion, migration, and invasion. Additionally, we performed Western blot to assess the expression of PI3K/Akt/mTOR-related proteins in PSAT1-knockdown cells. Results: The percentages of PSAT1-positive cells in pancreatic cancer and adjacent non-tumor tissues were 69.4% (68/98) and 5.0% (5/98), respectively, indicating a significantly higher expression of PSAT1 in pancreatic cancer tissues com-pared to adjacent non-tumor tissues (P<0.05). The increased expression of PSAT1 in pancreatic cancer tissues correlated with lymph node metastasis and TNM stage. Kaplan-Meier analysis suggested that a high expression of PSAT1 correlated with a poor OS and DFS compared to a low expression of PSAT1 (P<0.05). Cox regression analysis showed that the expression of PSAT1 is an independent prog-nostic marker for OS and DFS in pancreatic cancer patients (P<0.05, all). Transient transfection of BxPC-3 and SW1990 cells with PSAT1-siRNA markedly reduced the cell proliferation, migration, and invasion abilities of these cells compared to transfection with NC-siRNA (P<0.05). Knockdown of PSAT1 in pancreatic cancer cells also inhibited the expression of p-Akt and p-mTOR (P<0.05). Conclusions: The expression of PSAT1 increases in human pancreatic cancer tissues and cell lines. Additionally, PSAT1 regulates cell proliferation and in-vasion through the PI3K/Akt/mTOR pathway.

Hederacolchiside A1 suppresses proliferation of tumor cells by inducing apoptosis through modulating PI3K/Akt/mTOR signaling pathway / 中草药·英文版

Objective: Hederacolchiside A1, exhibits cytostatic and cytotoxic activity against various cancer cells in vitro, however, the mechanism is not well understood. Methods: In this study, Hederacolchiside A1 from Pulsatilla chinensis was isolated and tested its anticancer activity and mechanism. Hederacolchiside A1 could inhibit proliferation of A549, SMMC-7721, BEL-7402, and MCF-7 cells by MTT assay. Investigations of apoptosis of treated cancer cells were identified in hederacolchiside A1 by flow cytometric analysis of annexin V expression. Results: Based on the results of western blotting and JC-1 staining, hederacolchiside A1 reduced the mitochondrial membrane potential and Bcl-2 protein levels, whereas cleaved caspase-3 was higher. Furthermore, hederacolchiside A1 effectively inhibited the phosphorylations of phosphatidylinositol 3 kinase (PI3K), protein kinase B (Akt), and mammalian target of rapamycin (mTOR). In vivo study showed that hederacolchiside A1 (3.0, 4.5, and 6.0 mg/kg, ip) could significantly inhibit the weight of tumor in an H22 xenograft model. Similar inhibitory activities were observed when the compound (3.25, 7.5, and 15.0 mg/kg, ig) was tested in nude mice xenograft tumor models using human breast carcinoma MCF-7 cells. Conclusion: These data indicated that hederacolchiside A1 suppressed the proliferation of human tumor cells by inducing apoptosis through modulating the PI3K/Akt/mTOR signaling pathway.

Effects of Xinfeng Capsule on synovial angiogenesis based PI3K/AKT/mTOR pathway,HIF-1α,ES in rats model of adjuvant arthritis / 中国免疫学杂志

Objective:To observe the effect of Xinfeng Capsule (XFC) on synovial membrane PI3K/AKT/mTOR pathway in adjuvant arthritis (AA) rats.Methods:Rats were randomly divided into normal control group,model control group,methotrexate group,tripterygium glycoside group and XFC group.The expressions of IL-6,IL-10,HIF-1α and VEGF-A were detected by immunohistochemical method,and the expressions of PI3K,AKT1 and p-AKT1 were detected by immunoblotting.Results:Expression of AKT1,mTOR and ES proteins in synovial blood vessels.The expression of IL-6,VEGF,HIF-1α and synovial membrane PI3K,AKT1,p-AKT1,mTOR,ES in the model group were significantly higher than those in the control group The expression of PI3K,p-AKT1,mTOR,ES in synovial membrane decreased,and IL-10 in serum increased in XFC group,and the expression of HIF-1α,IL-6 and VEGF-A were decreased.Conclusion:XFC can improve angiogenesis in AA synovium by regulating PI3K/AKT/mTOR signaling pathway,HIF-1α and ES expression.

Effect of total flavonoids in Scutellaria barbata in mediating autophagy in tumor cells via PI3K/AKT/mTOR pathway / 中国中药杂志

To investigate the effect of the total flavonoids in Scutellaria barbata(TF-SB) against autophagy in tumor cells in vivo, and further determine whether the mechanism is correlated with the PI3K/AKT/mTOR pathway, which lead to autophagy-induced tumor cell death. Melanoma-bearing mice were prepared and divided into control group, rapamycin group (Rap 1.5 mg•kg⁻¹), and high, middle and low-dose TF-SB (200, 100, 50 mg•kg⁻¹) groups. The groups were given drugs once a day for successively 11 days. The inhibitory effect of TF-SB on the growth of melanoma was determined by measuring tumor volume and tumor inhibition rate. TUNEL method was used to detect the apoptosis of tumor cells to further verify the antitumor activity of TF-SB. The protein expressions of LC3-Ⅰ and LC3-Ⅱ were detected by Western blot, and the relative expression of LC3-Ⅱ was calculated based on LC3-Ⅱ/LC3-Ⅰ. In addition, the effect of TF-SB on autophagy of tumor cells, the underlying molecular mechanism of TF-SB in inducing autophagy and PI3K/AKT/mTOR pathway marker protein phosphorylation were also studied. According to the results, TF-SB effectively inhibited melanoma growth in mice, reduced tumor volume, increased the tumor inhibition rate, and significantly increased tumor cell apoptosis index and the ratio of LC3-Ⅱ/LC3-I (P<0.05, P<0.01 or P<0.001). The protein expressions of p-PI3K, p-AKT and p-mTOR were also suppressed dramatically compared with those in control group (P<0.05, P<0.01 or P<0.001). In conclusion, the total flavonoids in S. barbata could inhibit the growth of melanoma in vivo by inducing autophagy and apoptosis of tumor cells, which may be correlated with suppression of PI3K/AKT/mTOR pathway.

Effect of hydrogen sulfide on aβ25-35 -induced autophagy in mouse neuro-2a cells / 第二军医大学学报

Objective To investigate thêoeffect of exogenous hydrogen sulfide on Aβ25-35-induced autophagy in mouse Neuro-2a cells and the underlying mechanism. Methods The Neuro-2a cells were randomly divided into control group. Aβ25-35 treatment group. Aβ25-35 + NaHS group. Aβ25-35 n3-MA (3-methyl adenine) group. NaHS group and Aβ25-35 + NaHS + LY294002 group. In Aβ25-35+ NaHS group and Aβ25-35 +3-MA group. NaHS and 3-MA were used for pretreatment for 2 h. which was followed by 24 h co-incubation with Aβ5-35; in Aβ25-35 + NaHS+LY294002 group. after pretreatment with NaHS. LY294002 was administered for 0. 5 h before Aβ25-35 was given. MTT assay was used to detect the viability of Neuro-2a cells. The expression of the autophagy related proteins including Beclin-1. LC3 and P62was measured by Western blotting analysis. Immunofluorescence was used to detect the expression and distribution of LC3. The formation ofautophagosomes was determined by transmission electron microscopy (TEM). Results (1) Compared with control group. Aβ25-35 group had significantly declined cell viability (P<0. 05). significantly increased expression of Beclin-1 and LC3H and decreased P62 expression (P<0. 05). accompanied by increased autophagsomes under fluorescencemicroscope and electron microscope. (2) Compared with Aβ25-35 group. the cell viability was significantly increased in Aβ25-35 + 3-MA and Aβ25-35 +NaHS groups (P< 0. 05). and expression of Beclin-1 and LC3H was decreased and expression of P62 was increased (P<0. 05). accompanied by reduced autophagosomes. (3) The expression of p-Akt and p-mTOR in Aβ25-35 +NaHS group was significantly higher than that in Aβ25-35 group (P<0. 05); however. the expression of p-Akt and p-mTOR was significantly reduced after given the specific PI3K/Akt pathway inhibitor LY294002 compared with Aβ25-35 +NaHS group (P<0. 05). Conclusion The exogenous H2S can protect against Aβ25-35 -induced cytotoxicity. which is probably related to the activation of PI3K/Akt/mTOR pathways and the inhibition of Aβ25-35 -induced cell autophagy.

Effects of 1,25-(OH) 2 D3 on Ca2+concentration and PI3K/AKT/mTOR pathway in type Ⅱalveolar epi-thelial cells of rat with pulmonary fibrosis / 医学研究生学报

Objective Idiopathic pulmonary fibrosis ( IPF) is a chronic inflammatory disease with unknown etiology and is lack of effective therapy. The aim of this study is to explore the function of Ca2+ and PI3K/AKT/mTOR pathway in the pathogenesis of IPF, and the impact of 1,25?( OH) 2 D3 on Ca2+ and PI3K/AKT/mTOR pathway in type Ⅱalveolar epithelial cells of rat with IPF. Methods 150 male SD rats were randomly divided into 2 groups: prevention group ( control groupⅠ, model groupⅠ, medication groupⅠ) and treatment group ( control groupⅡ, model groupⅡ, medication groupⅡ) . The tracheal exposure surgery was operated in control groupⅠ/Ⅱ, and then 200μL sterile physiological saline was administered by intraperitoneal injection of each rats 2 days and 14 days after surgery, separately. Bleomycin(BLM)(5 mg/kg) was in?jected into the trachea of model groupⅠ/Ⅱ, and then vitamin D3 solvent(0.1%ethanol and 99.9%glycol propylene, 1μL/g) was ad?ministered by intraperitoneal injection 2 days and 14 days after surger?y, separately. Bleomycin( BLM) ( 5 mg/kg) was injected into the tra?chea of medication groupⅠ/Ⅱ, and then 1,25?( OH) 2 D3( 2μg/kg) was administered by intraperitoneal injection 2 days and 14 days after surgery, separately. IPF model was built by injecting Bleomycin into the trachea of rats, 1,25?(OH)2D3(2μg/kg) was used to prevent and treat IPF by intraperitoneal injection in medication group. The hydroxyproline content of lung tissue in each group was measured, type Ⅱalveolar epithelial cells were separated from lung tissue and labeled with Fluo?3AM, then concentration of Ca2+ was detected by Laser scanning confocal microscope. The mRNA levels of PI3K, AKT and mTOR in the typeⅡalveolar epithelial cells were tested by RT?PCR. Results Compared with control groupⅠ/Ⅱ at each time point, hydroxyproline content of lung tissue, Ca2+ concentration and expression of PI3K, AKT and mTOR in typeⅡalveolar epithelial cells in model groupⅠ/Ⅱand medication groupⅠ/Ⅱwere sig?nificantly raised( P<0.05 or P<0.01) , but these were significantly reduced in medication groupⅠ/Ⅱcompared with model groupⅠ/Ⅱ( P<0.05 or P<0.01) . Correlation analysis showed that there is significant positive correlation between Ca2+ concentration and mRNA expression levels of PI3K, AKT and mTOR in model groupⅠ/Ⅱ(r=0.5988, r=0.6230, r=0.6603,P<0.01)and medication groupⅠ/Ⅱ( r=0.701 2, r=0.632 3,r=0.740 3,P<0.01) . Conclusion The PI3K/AKT/mTOR pathway plays an important role in devel?opment of IPF. 1,25?( OH) 2 D3 is able to reduce Ca2+concentration in typeⅡalveolar epithelial cells and inhibit the PI3K/AKT/mTOR pathway, and then inhibit the development of IPF.

Research progress of small molecule inhibitors targeting PI3K-Akt-mTOR pathway / 中国生化药物杂志

Aberrant activation of the phosphatidylinositol 3-kinase(PI3K)-protein kinase B(PKB,Akt)-mammalian target of rapamycin(mTOR) pathway is commonly observed in human cancer and is critical for cell survival, proliferation and differentiation.A variety of small molecule inhibitors targeting PI3K-Akt-mTOR pathway are under clinical studies.This review will summarize the recent studies in terms of the PI3K-Akt-mTOR signaling pathway and cancer,research progress of the antitumor activity possessed by PI3K-Akt-mTOR inhibitors,as well as the recent research in the related field conducted by our group.

Research progress on Chinese materia medica in overcoming EGFR-TKIs acquired drugs resistance through PI3K/Akt/mTOR pathway / 中草药

With in-depth study of the pathogenesis of lung cancer and its biological behaviour, molecular targeted therapy, especially the epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) made a breakthrough in non-small cell lung cancer treatment. However, the majority of patients can produce the resistance to EGFR-TKIs after 6 to 12 months treatment, which called acquired drug resistance. PI3K/Akt/mTOR pathway plays an important role in the development of non-small cell lung cancer, this pathway in cancer treatment has been widely studied. Chinese materia medica (CMM) in overcoming EGFR-TKIs resistance has a bright future, In this review, we show the developments on CMM in overcoming EGFR-TKIs acquired drugs resistance through PI3K/Akt/mTOR pathway.

The research development of PI3K/AKT/mTOR signaling pathway in Her-2 resistance for the treat-ment of breast cancer / 实用肿瘤学杂志

Her-2 targeting therapy is an important part of the treatment for breast cancer with Her -2 overexpression.However,the therapeutic effectiveness is largely influenced by Her -2 resistance.Previous studies have confirmed that PI3K/AKT/mTOR signaling pathway is activated during the process of drug resistance to Her-2-targeting therapy.Therefore,investigating the PI3K/AKT/mTOR signaling pathway and drugs -targeted pathway are important for the treatment of breast cancer .

Improvement effects of astragaloside Ⅳ on myocardial focal ischemia-reperfusion inj ury and its influence in PI3K/Akt/mTOR signaling pathway / 吉林大学学报(医学版)

Objective To observe the improvement effects of astragaloside Ⅳ (AS-Ⅳ)on the myocardial focal ischemia-reperfusion (I/R)injury and its influence in PI3K/Akt/mTOR pathway,and to clarify the protective effect of AS-Ⅳ on myocardial I/R inj ury and the possible mechanisms.Methods The left main coronary arteries of 60 Sprague-Dawley rats were occluded for 30 min followed by a 120-min reperfusion to induce I/R model.The rats with I/R inj ury were randomly divided into model group (normal saline), AS-Ⅳ group (intravenous inj ection of 10 mg·kg-1 AS-Ⅳ 5 min before reperfusion),PI3K inhibitor Wortmannin (WOR)group (intravenous injection of 0.6 mg·kg-1 WOR 10 min before reperfusion)and AS-Ⅳ+WOR group (intravenous injection of 10 mg·kg-1 AS-Ⅳ and 0.6 mg·kg-1 WOR 5 and 10 min before reperfusion,respectively).15 age-matched SD rats were chosen as control group.The heart mass,degrees of infarction and ischemia and cardiac function ,including left ventricular systolic mean pressure (LVSP),end-diastolic pressure (LVEDP),fractional shortening (FS)and ej ection fraction (EF),of the rats in all groups were analyzed. Western blotting method was used to measure the phosphorylation levels of Akt and mTOR(p-Akt and p-mTOR).The specific fluorescent probe DHE staining was employed to detect the myocardial reactive oxygen species levels. Results Compared with control group, the degrees of infarction and ischemia, LVEDP, myocardial levels of p-Akt/Akt, p-mTOR/mTOR and reactive oxygen species levels of the rats were increased (P<0.05).and the levels of LVSP,FS and EF were decreased in model group (P<0.05). Compared with the model group,the degrees of infarction and ischemia,LVEDP and reactive oxygen species level were decreased (P<0.05),while the levels of p-Akt/Akt,p-mTOR/mTOR LVSP, FS and EF of all rats in AS-Ⅳ group were increased (P<0.05).Compared with AS-Ⅳ group,the degrees of infarction and ischemia,LVEDP and reactive oxygen species levels of the rats in WOR group and AS-Ⅳ+WOR group were increased (P<0.05 ), and the myocardial p-Akt/Akt, p-mTOR/mTOR and LVSP, FS, EF were decreased (P<0.05).Conclusion AS-Ⅳ has improvement effect on myocardial I/R injury.AS-Ⅳ can reduce the extent of myocardial infarction and oxidative stress and improve the heart function,and its possible mechanism may be related to activating PI3K/Akt/mTOR signaling pathway.