AIM： To study the expression level and localization of insulin-like growth factor -Ⅰ receptor （IGF-IR） in HepG2 cells and Chang liver cells, and to observe the effect of anti-IGF-IR monoclonal antibody （αIR3） o...AIM： To study the expression level and localization of insulin-like growth factor -Ⅰ receptor （IGF-IR） in HepG2 cells and Chang liver cells, and to observe the effect of anti-IGF-IR monoclonal antibody （αIR3） on the growth of HepG2 cells. METHODS： The expression of IGF-IR in HepG2 cells and Chang liver cells was detected by immunohistochemistry. The influences of αIR3 on proliferation and apoptosis were examined by the 3- （4, 5-dimethylthiazol-2-yl）-2, 5- diphenyltetrazolium bromide （MTT） assay and electron microscopy, respectively. Flow cytometry （FCM） was applied for the analysis of cell cycle and apoptosis was observed under electron microscope. RESULTS： IGF-IR was located in the membranes of both HepG2 and Chang liver cell lines, and the expression level of IGF-IR was higher in HepG2 cells than in Chang liver cells. Treated with 0.1 μg/mL αIR3 for 48 h in vitro, the cell growth index （GI） of HepG2 cells was significantly higher than that of control （103.41% ys 100%, P 〈 0.01）. However, the αIR3 for 24 h at final concentration of 4.0 μg/mL made the GI of HepG2 cells lower than that of control （93.37% vs 100%, P 〈 0.01）. Compared with control, treated with αIR3 for 48 h at final concentrations ranging from 2.0 μg/mL to 4.0 μg/mL markedly reduced the GIs of HepG2 cells （97.63%, 97.16%, 95.13%, 92.53% vs 100%, P 〈 0.05 or P 〈 0.01）, treated with αIR3 for 72 h at final concentrations ranging from 0.2 μg/mL to 4.0 μg/mL decreased the GIs of HepG2 cells obviously （95%, 91.63%, 90.77%, 89.84%, 88.51% vs 100%, P 〈 0.01）, and treated with αIR3 for 96 h at final concentrations ranging from 0.5 μg/mL to 4.0 μg/mL made GIs of HepG2 cells lower significantly （88.86%, 83.97%, 79.81%, 77.24%, 70.51% vs 100%, P 〈 0.05or P 〈 0.01）. Moreover, treated with αIR3 from 24 h to 96 h at final concentrations ranging from 0.2 μg/mL to 4.0 μg/mL reduced the GI of HepG2 cells from 97.63% to 70.51% in a dose- and time-dependent manner. Also, αIR3 treatment for 72 h at final concentration from 0.5 μg/mL to 2.0 μg/mL increased the proportion of G0/G1 phase cells（61.73%, 67.1%, 83.7%,76.87% vs 44.47%, P 〈 0.01） and significantly decreased that of S phase cells（28.63%, 25.13%, 15.63%, 23.13% vs 53.17%, P 〈 0.01）, in contrast to the proportion of G2/M phase cells. The apoptotic rates of HepG2 cells were increased more than that of control （7.83%, 16.13%, 21.1%, 37.73% vs 4.13%, P 〈 0.01）. CONCLUSION： The malignant cell phenotype of human hepatocarcinoma cell is related to overexpression of IGF- IR. The blockage of IGF-IR with αIR3 may contribute to the inhibition of proliferation and induction of apoptosis in HepG2 cells.展开更多
Background and Aims:The results of basic research implicate the vascular endothelial growth factor(VEGF)family as a potential target of hepatopulmonary syndrome(HPS).However,the negative results of anti-angiogenetic t...Background and Aims:The results of basic research implicate the vascular endothelial growth factor(VEGF)family as a potential target of hepatopulmonary syndrome(HPS).However,the negative results of anti-angiogenetic therapy in clinical studies have highlighted the need for markers for HPS.Therefore,we aimed to determine whether VEGF family members and their receptors can be potential biomarkers for HPS through clinical and experimental studies.Methods:Clinically,patients with chronic liver disease from two medical centers were enrolled and examined for HPS.Patients were divided into HPS,intrapulmonary vascular dilation[positive contrast-enhanced echocardiography(CEE)and normal oxygenation]and CEE-negative groups.Baseline information and perioperative clinical data were compared between HPS and non-HPS patients.Serum levels of VEGF family members and their receptors were measured.In parallel,HPS rats were established by common bile duct ligation.Liver,lung and serum samples were collected for the evaluation of pathophysiologic changes,as well as the expression levels of the above factors.Results:In HPS rats,all VEGF family members and their receptors underwent significant changes;however,only soluble VEGFR1(sFlt-1)and the sFlt-1/placental growth factor(PLGF)ratio were changed in almost the same manner as those in HPS patients.Furthermore,through feature selection and internal and external validation,sFlt-1 and the sFlt-1/PLGF ratio were identified as the most important variables to distinguish HPS from non-HPS patients.Conclusions:Our results from animal and human studies indicate that sFlt-1 and the sFlt-1/PLGF ratio in serum are potential markers for HPS.展开更多
Intraductal papilloma of the breast is relatively common,accounting for 5.3%of all benign breast diseases.[1]However,relapse readily occurs with pathological changes such as atypical ductal hyperplasia(ADH)and cancera...Intraductal papilloma of the breast is relatively common,accounting for 5.3%of all benign breast diseases.[1]However,relapse readily occurs with pathological changes such as atypical ductal hyperplasia(ADH)and canceration,and the rate of underestimation of biopsy diagnosis is high.[2,3]Therefore,specific considerations are required for the diagnosis and treatment methods of intraductal papilloma,and some clinical problems with these methods remain controversial.To standardize the diagnosis and treatment of intraductal papilloma of the breast and provide a reference for the clinical work of breast specialists,Chinese Society of Breast Surgery has determined the key clinical issues of the clinical practice guidelines for intraductal papilloma of the breast through a literature search and expert discussion.The relevant evidence was evaluated with reference to the Grading of Recommendations Assessment,Development and Evaluation(GRADE)system,and the Clinical Practice Guidelines for Intraductal Papilloma:Chinese Society of Breast Surgery(CSBrS)practice guidelines 2021 was formulated to provide a reference for the clinical work of breast surgeons in China.展开更多
Background:Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far.Petasin,a natural product found in plants of the genus Petasites,has been reported to possess anticance...Background:Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far.Petasin,a natural product found in plants of the genus Petasites,has been reported to possess anticancer activity.The present study aimed to investigate the anticolon cancer activity of petasin both in vitro and in vivo.The molecular mechanism of petasin was also further explored.Methods:Caco-2,LoVo,SW-620,and HT-29 cell lines were used to detect the inhibitory effect of petasin on colon cancer proliferation.Cell viability was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay.Cell apoptosis was analyzed by flow cytometry.Hoechst 33258 staining was used to visualize morphological changes.Cell migration was assessed using a wound-healing migration assay,and cell invasion was investigated using Transwell chambers.Western blotting assays were employed to evaluate the expression levels of proteins in the protein kinase B/mammalian target of rapamycin (Akt/mTOR) signaling pathway.Finally,in vivo activity of petasin was evaluated using the SW-620 subcutaneous tumor model established in Balb/c nude mice.Twelve rats were randomly divided into control group and 10 mg/kg petasin group.The tumor volume was calculated every 7 days for 28 days.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was performed to assess the apoptotic effect of petasin.Differences between two groups were assessed by analysis of independent-sample t tests.Results:Petasin significantly inhibited the proliferation of human colon carcinoma cell lines,induced apoptosis,and suppressed migration and invasion in SW-620 cells.Western blotting results showed that petasin decreased the phosphorylation of Akt (1.01 ± 0.16 vs.0.74 ± 0.06,P = 0.042),mTOR (0.71 ± 0.12 vs.0.32 ± 0.11,P = 0.013),and P70S6K (1.23 ± 0.21 vs.0.85 ± 0.14,P = 0.008),elevated the expression of caspase-3 (0.41 ± 0.09 vs.0.74 ± 0.12,P = 0.018) and caspase-9 (1.10 ± 0.27 vs.1.98 ± 0.22,P = 0.009),decreased the Bcl-2 protein (2.75 ± 0.47 vs.1.51 ± 0.36,P = 0.008),downregulated the expression of matrix metalloproteinase (MMP)-3 (1.51 ± 0.31 vs.0.82 ± 0.11,P = 0.021) and MMP-9 (1.56 ± 0.32 vs.0.94 ± 0.15,P = 0.039) in SW-620 cell.In vivo,10 mg/kg petasin inhibited tumor growth in Balb/c nude mice (924.18 ± 101.23 vs.577.67 ± 75.12 mm3 at day 28,P = 0.001) and induced apoptosis (3.6 ± 0.7% vs.36.0 ± 4.9%,P = 0.001) in tumor tissues.Conclusions:Petasin inhibits the proliferation of colon cancer SW-620 cells via inactivating the Akt/mTOR pathway.Our findings suggest petasin as a potential candidate for colon cancer therapy.展开更多
基金Supported by the Gansu Province's Natural Science Fund, No.ZS021-A25-079-Y
文摘AIM： To study the expression level and localization of insulin-like growth factor -Ⅰ receptor （IGF-IR） in HepG2 cells and Chang liver cells, and to observe the effect of anti-IGF-IR monoclonal antibody （αIR3） on the growth of HepG2 cells. METHODS： The expression of IGF-IR in HepG2 cells and Chang liver cells was detected by immunohistochemistry. The influences of αIR3 on proliferation and apoptosis were examined by the 3- （4, 5-dimethylthiazol-2-yl）-2, 5- diphenyltetrazolium bromide （MTT） assay and electron microscopy, respectively. Flow cytometry （FCM） was applied for the analysis of cell cycle and apoptosis was observed under electron microscope. RESULTS： IGF-IR was located in the membranes of both HepG2 and Chang liver cell lines, and the expression level of IGF-IR was higher in HepG2 cells than in Chang liver cells. Treated with 0.1 μg/mL αIR3 for 48 h in vitro, the cell growth index （GI） of HepG2 cells was significantly higher than that of control （103.41% ys 100%, P 〈 0.01）. However, the αIR3 for 24 h at final concentration of 4.0 μg/mL made the GI of HepG2 cells lower than that of control （93.37% vs 100%, P 〈 0.01）. Compared with control, treated with αIR3 for 48 h at final concentrations ranging from 2.0 μg/mL to 4.0 μg/mL markedly reduced the GIs of HepG2 cells （97.63%, 97.16%, 95.13%, 92.53% vs 100%, P 〈 0.05 or P 〈 0.01）, treated with αIR3 for 72 h at final concentrations ranging from 0.2 μg/mL to 4.0 μg/mL decreased the GIs of HepG2 cells obviously （95%, 91.63%, 90.77%, 89.84%, 88.51% vs 100%, P 〈 0.01）, and treated with αIR3 for 96 h at final concentrations ranging from 0.5 μg/mL to 4.0 μg/mL made GIs of HepG2 cells lower significantly （88.86%, 83.97%, 79.81%, 77.24%, 70.51% vs 100%, P 〈 0.05or P 〈 0.01）. Moreover, treated with αIR3 from 24 h to 96 h at final concentrations ranging from 0.2 μg/mL to 4.0 μg/mL reduced the GI of HepG2 cells from 97.63% to 70.51% in a dose- and time-dependent manner. Also, αIR3 treatment for 72 h at final concentration from 0.5 μg/mL to 2.0 μg/mL increased the proportion of G0/G1 phase cells（61.73%, 67.1%, 83.7%,76.87% vs 44.47%, P 〈 0.01） and significantly decreased that of S phase cells（28.63%, 25.13%, 15.63%, 23.13% vs 53.17%, P 〈 0.01）, in contrast to the proportion of G2/M phase cells. The apoptotic rates of HepG2 cells were increased more than that of control （7.83%, 16.13%, 21.1%, 37.73% vs 4.13%, P 〈 0.01）. CONCLUSION： The malignant cell phenotype of human hepatocarcinoma cell is related to overexpression of IGF- IR. The blockage of IGF-IR with αIR3 may contribute to the inhibition of proliferation and induction of apoptosis in HepG2 cells.
基金supported by National Science Foundation of China(No.82070630 from Bin Yi,82100658 from Yu-jie Li and 82170634 from Peng Li)National Key R&D Program of China(No.2018YFC0116702 from Bin Yi)+2 种基金Special support for Chongqing postdoctoral research project in 2020 from Yujie Li,Sichuan science and technology department research projects(2019YFS0221 from Peng Li)Chongqing Science and health joint medical research project(2020FYYX076,from Bin Yi)special support project for improving scientific and technological innovation ability of undergraduate(2021XBK19 from Xian-feng Wu).
文摘Background and Aims:The results of basic research implicate the vascular endothelial growth factor(VEGF)family as a potential target of hepatopulmonary syndrome(HPS).However,the negative results of anti-angiogenetic therapy in clinical studies have highlighted the need for markers for HPS.Therefore,we aimed to determine whether VEGF family members and their receptors can be potential biomarkers for HPS through clinical and experimental studies.Methods:Clinically,patients with chronic liver disease from two medical centers were enrolled and examined for HPS.Patients were divided into HPS,intrapulmonary vascular dilation[positive contrast-enhanced echocardiography(CEE)and normal oxygenation]and CEE-negative groups.Baseline information and perioperative clinical data were compared between HPS and non-HPS patients.Serum levels of VEGF family members and their receptors were measured.In parallel,HPS rats were established by common bile duct ligation.Liver,lung and serum samples were collected for the evaluation of pathophysiologic changes,as well as the expression levels of the above factors.Results:In HPS rats,all VEGF family members and their receptors underwent significant changes;however,only soluble VEGFR1(sFlt-1)and the sFlt-1/placental growth factor(PLGF)ratio were changed in almost the same manner as those in HPS patients.Furthermore,through feature selection and internal and external validation,sFlt-1 and the sFlt-1/PLGF ratio were identified as the most important variables to distinguish HPS from non-HPS patients.Conclusions:Our results from animal and human studies indicate that sFlt-1 and the sFlt-1/PLGF ratio in serum are potential markers for HPS.
文摘Intraductal papilloma of the breast is relatively common,accounting for 5.3%of all benign breast diseases.[1]However,relapse readily occurs with pathological changes such as atypical ductal hyperplasia(ADH)and canceration,and the rate of underestimation of biopsy diagnosis is high.[2,3]Therefore,specific considerations are required for the diagnosis and treatment methods of intraductal papilloma,and some clinical problems with these methods remain controversial.To standardize the diagnosis and treatment of intraductal papilloma of the breast and provide a reference for the clinical work of breast specialists,Chinese Society of Breast Surgery has determined the key clinical issues of the clinical practice guidelines for intraductal papilloma of the breast through a literature search and expert discussion.The relevant evidence was evaluated with reference to the Grading of Recommendations Assessment,Development and Evaluation(GRADE)system,and the Clinical Practice Guidelines for Intraductal Papilloma:Chinese Society of Breast Surgery(CSBrS)practice guidelines 2021 was formulated to provide a reference for the clinical work of breast surgeons in China.
文摘Background:Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far.Petasin,a natural product found in plants of the genus Petasites,has been reported to possess anticancer activity.The present study aimed to investigate the anticolon cancer activity of petasin both in vitro and in vivo.The molecular mechanism of petasin was also further explored.Methods:Caco-2,LoVo,SW-620,and HT-29 cell lines were used to detect the inhibitory effect of petasin on colon cancer proliferation.Cell viability was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay.Cell apoptosis was analyzed by flow cytometry.Hoechst 33258 staining was used to visualize morphological changes.Cell migration was assessed using a wound-healing migration assay,and cell invasion was investigated using Transwell chambers.Western blotting assays were employed to evaluate the expression levels of proteins in the protein kinase B/mammalian target of rapamycin (Akt/mTOR) signaling pathway.Finally,in vivo activity of petasin was evaluated using the SW-620 subcutaneous tumor model established in Balb/c nude mice.Twelve rats were randomly divided into control group and 10 mg/kg petasin group.The tumor volume was calculated every 7 days for 28 days.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was performed to assess the apoptotic effect of petasin.Differences between two groups were assessed by analysis of independent-sample t tests.Results:Petasin significantly inhibited the proliferation of human colon carcinoma cell lines,induced apoptosis,and suppressed migration and invasion in SW-620 cells.Western blotting results showed that petasin decreased the phosphorylation of Akt (1.01 ± 0.16 vs.0.74 ± 0.06,P = 0.042),mTOR (0.71 ± 0.12 vs.0.32 ± 0.11,P = 0.013),and P70S6K (1.23 ± 0.21 vs.0.85 ± 0.14,P = 0.008),elevated the expression of caspase-3 (0.41 ± 0.09 vs.0.74 ± 0.12,P = 0.018) and caspase-9 (1.10 ± 0.27 vs.1.98 ± 0.22,P = 0.009),decreased the Bcl-2 protein (2.75 ± 0.47 vs.1.51 ± 0.36,P = 0.008),downregulated the expression of matrix metalloproteinase (MMP)-3 (1.51 ± 0.31 vs.0.82 ± 0.11,P = 0.021) and MMP-9 (1.56 ± 0.32 vs.0.94 ± 0.15,P = 0.039) in SW-620 cell.In vivo,10 mg/kg petasin inhibited tumor growth in Balb/c nude mice (924.18 ± 101.23 vs.577.67 ± 75.12 mm3 at day 28,P = 0.001) and induced apoptosis (3.6 ± 0.7% vs.36.0 ± 4.9%,P = 0.001) in tumor tissues.Conclusions:Petasin inhibits the proliferation of colon cancer SW-620 cells via inactivating the Akt/mTOR pathway.Our findings suggest petasin as a potential candidate for colon cancer therapy.
