scholarly journals The Role of Apatinib Combined with Paclitaxel (aluminum binding type) in Platinum-resistant Ovarian Cancer

2020 ◽  
Author(s):  
Hong Zhao ◽  
Rong Li ◽  
Xiaoyan Wang ◽  
Xin Lu ◽  
Min Hu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Western Blot ◽  
Nude Mice ◽  
Scratch Test ◽  
Control Group ◽  
Related Protein ◽  
Drug Intervention ◽  
Platinum Resistant ◽  
Xenograft Models ◽  
Apoptosis Related Protein

Abstract Objective: To assess the anti-tumor activity and side effects of different dosages of paclitaxel (albumin binding type) (hereinafter referred to as nab-P) combined with Apatinib (hereinafter referred to as AP) in platinum-resistant ovarian cancer cell line and xenograft models.Methods: SKOV-3/DDP cell line was selected as the research object in cytology experiment. Firstly, we divided it into three groups for experiments to explore the individual effects of nab-P and AP. a): Control group, blank control, no drug intervention; b): nab-P group, nab-P 40μmol/l; c): AP group, AP 50μmol/l (Drug doses were IC-50 values that detected by MTT assay). Apoptosis related protein(Bax, bcl-2), vascular related protein(p-VEGFR-2), invasion related protein(MMP-2) expression were detected by Western blot and Cellular immunofluorescence, the invasion ability of tumor cells were detected by Transwell and Cell scratch test. Based on these dates, secondly, establishing different doses of nab-P combined with Ap to explore the curative effect of combination therapy. a): Control group, blank control, no drug intervention; b): Group-1, nab-P 5μmol/l + AP 10μmol/l, c): Group-2, nab-P 4.5μmol/l + AP 10μmol/l, d): Group-3, nab-P 4μmol/l + AP 10μmol/l, e): nab-P group, nab-P 5μmol/l, f): AP group, AP 10μmol/l (MTT assay). The combination index was analyzed by Compusyn software, Western blot, Immunofluescence, Transwell and Cell scratch test also were also chose to observe of inhibition effect. Thirdly, we used xenograft models to verify the results of cytological experiments. Tumor-forming BALB/c female nude mice were randomly divided into 4 groups, a): Control group, no drug intervention, only saline injection, b): nab-P 20mg/kg + AP 150mg/kg , c): nab-P 18mg/kg + AP 150mg/kg, d): nab-P 16mg/kg + AP 150mg/kg(The doses were guided by the pharmaceutical manufacturers). The tumor growth curve was analyzed during the experiment. And the apoptosis related protein (Bax, bcl-2), angiogenesis related protein (CD31, p-VEGFR-2) and invasion related protein(MMP-2) were observed by Western blot, Immunofluescence and Immunohistochemistry to analysis the ant-tumor effects. The quality of life in nude mice were observed to analysed the drug-induced side effects.Result: In the separate medication section, (1) The IC-50 value of nab-P was 45.53±4.06 μmol/l, while the AP was 50.66±4.96 umol/L (48h). (2) The expressions of bcl-2 (nab-P group, AP group), p-VEGFR-2 (AP group), MMP-2(nab-P group, AP group) were higher than Control group, while Bax(nab-P group, AP group) lower (P<0.01). (3) The cell invasive ability was decreased after the nab-P and AP intervation (P<0.01). In the combination medication section, (1) Compusyn showed the Combination index (Cl) were all below 1 (Cl<1), that means nab-P and AP are synergism. (2) The combination IC-50 value was nab-P 5.28 μmol/l+AP 10.56 μmol/l (48h). (3) In the detection of related protein expression, the combination of drugs can improve the anti-tumor effect, otherwise, after combined with AP, when nab-P were reduced dose in proper quantity, there were no obvious different in drug effect. (4) After reducing the doses of nab-P, the average food intake of nude mice increased from 4.50 g±0.17 to 5.55 g±0.13, and the one-hour activity increased from 6.11min ±0.16 to 6.34 min ±0.13.Conclusion: nab-P, a chemotherapeutic agent, can play an anti-tumor role in platinum-resistant ovarian cancer, but it can cause adverse effects that increase with dose. When combined with AP, the two drugs have synergistic effect, which can improve the anti-tumor effects of single drug. In addition, when combined with AP, the doses of nab-P can be appropriately reduced under the standard of recommended to reduce the toxicity of chemotherapy drugs, without affecting the anti-tumor effect.

scholarly journals The role of apatinib combined with paclitaxel (aluminum binding type) in platinum-resistant ovarian cancer

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Hong Zhao ◽  
Rong Li ◽  
Xiaoyan Wang ◽  
Xin Lu ◽  
Min Hu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Western Blot ◽  
Nude Mice ◽  
Scratch Test ◽  
Control Group ◽  
Related Protein ◽  
Drug Intervention ◽  
Platinum Resistant ◽  
Xenograft Models ◽  
Apoptosis Related Protein

Abstract Objective To assess the anti-tumor activity and side effects of different dosages of paclitaxel (albumin binding type) (hereinafter referred to as nab-P) combined with Apatinib (hereinafter referred to as AP) in platinum-resistant ovarian cancer cell line and xenograft models. Methods SKOV-3/DDP cell line was selected as the research object in cytology experiment. Firstly, we divided it into three groups for experiments to explore the individual effects of nab-P and AP. a): Control group, blank control, no drug intervention; b): nab-P group, nab-P 40 μmol/l; c): AP group, AP 50 μmol/l (Drug doses were IC-50 values that detected by MTT assay). Apoptosis related protein (Bax, bcl-2), vascular related protein(p-VEGFR-2), invasion related protein (MMP-2) expression were detected by Western blot and Cellular immunofluorescence, the invasion ability of tumor cells were detected by Transwell and Cell scratch test. Based on these dates, secondly, establishing different doses of nab-P combined with Ap to explore the curative effect of combination therapy. a): Control group, blank control, no drug intervention; b): Group-1, nab-P 5 μmol/l + AP 10 μmol/l, c): Group-2, nab-P 4.5 μmol/l + AP 10 μmol/l, d): Group-3, nab-P 4 μmol/l + AP 10 μmol/l, e): nab-P group, nab-P 5 μmol/l, f): AP group, AP 10 μmol/l (MTT assay). The combination index was analyzed by Compusyn software, Western blot, Immunofluescence, Transwell and Cell scratch test also were also chose to observe of inhibition effect. Thirdly, we used xenograft models to verify the results of cytological experiments. Tumor-forming BALB/c female nude mice were randomly divided into 4 groups, a): Control group, no drug intervention, only saline injection, b): nab-P 20 mg/kg + AP 150 mg/kg, c): nab-P 18 mg/kg + AP 150 mg/kg, d): nab-P 16 mg/kg + AP 150 mg/kg (The doses were guided by the pharmaceutical manufacturers). The tumor growth curve was analyzed during the experiment. And the apoptosis related protein (Bax, bcl-2), angiogenesis related protein (CD31, p-VEGFR-2) and invasion related protein (MMP-2) were observed by Western blot, Immunofluescence and Immunohistochemistry to analysis the ant-tumor effects. The quality of life in nude mice were observed to analysed the drug-induced side effects. Result In the separate medication section, (1) The IC-50 value of nab-P was 45.53 ± 4.06 μmol/l, while the AP was 50.66 ± 4.96 umol/L (48 h). (2) The expressions of bcl-2 (nab-P group, AP group), p-VEGFR-2 (AP group), MMP-2(nab-P group, AP group) were higher than Control group, while Bax (nab-P group, AP group) lower (P < 0.01). (3) The cell invasive ability was decreased after the nab-P and AP intervation (P < 0.01). In the combination medication section, (1) Compusyn showed the Combination index (Cl) were all below 1 (Cl < 1), that means nab-P and AP are synergism. (2) The combination IC-50 value was nab-P 5.28 μmol/l + AP 10.56 μmol/l (48 h). (3) In the detection of related protein expression, the combination of drugs can improve the anti-tumor effect, otherwise, after combined with AP, when nab-P were reduced dose in proper quantity, there were no obvious different in drug effect. (4) After reducing the doses of nab-P, the average food intake of nude mice increased from 4.50 g ± 0.17 to 5.55 g ± 0.13, and the one-hour activity increased from 6.11 min ±0.16 to 6.34 min ±0.13. Conclusion nab-P, a chemotherapeutic agent, can play an anti-tumor role in platinum-resistant ovarian cancer, but it can cause adverse effects that increase with dose. When combined with AP, the two drugs have synergistic effect, which can improve the anti-tumor effects of single drug. In addition, when combined with AP, the doses of nab-P can be appropriately reduced under the standard of recommended to reduce the toxicity of chemotherapy drugs, without affecting the anti-tumor effect.

scholarly journals The role of Apatinib combined with Paclitaxel (aluminum binding type) in drug-resistant ovarian cancer

2020 ◽  
Author(s):  
Hong Zhao ◽  
Rong Li ◽  
Xiaoyan Wang ◽  
Xin Lu ◽  
Min Hu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Western Blot ◽  
Tumor Model ◽  
Control Group ◽  
Side Reactions ◽  
Albumin Binding ◽  
Drug Resistant ◽  
Related Protein ◽  
Drug Intervention ◽  
Binding Type

Abstract Objective To assess the antitumor effects and side reactions of different dosages of Paclitaxel (albumin binding) combined with Apatinib in drug-resistant ovarian cancer cell line and xenotransplantation tumor model. Methods Conventional cell experiments were used to evaluate the effects of Apatinib and Paclitaxel (albumin binding), SKOV-3/DDP were selected as research object and divided into 3 groups for study, a): control group, no drug intervention; b): nab-P group, Paclitaxel (albumin binding type) 40 µmol/l; c): Apatinib group, Apatinib 50 µmol/l. The IC-50 value of the drug was detected by MTT test, apoptosis related protein(Bax, bcl-2), vascular related protein(p-VEGFR-2), invasion related protein(MMP-2) expression were detected by Western blot and Cellular immunofluorescence, the invasion and migration ability of tumor cells were detected by Transwell and Cell scratch test. Based on these dates, establishing different dosages of Paclitaxel (albumin binding type) combined with Apatinib, a): Control group, no drug intervention; b): Group-1, Paclitaxel (albumin binding type) 5 µmol/l + Apatinib 10 µmol/l, c): Group-2, Paclitaxel (albumin binding type) 4.5 µmol/l + Apatinib 10 µmol/l, d): Group-3, Paclitaxel (albumin binding type)4 µmol/l + Apatinib 10 µmol/l, the index of combined use was analyzed by Compusyn software, Western blot, Immunofluescence, Transwell, Cell scratch test also were chose to change of inhibition effect. On the other hand, we used xenograft tumor model to verify the results in vivo. BALB/c female nude mice were randomly divided into 4 groups, a): Control group, no drug intervention; b): Paclitaxel (albumin binding type) 20 mg/kg + Apatinib 150 mg/kg, c): Paclitaxel (albumin binding type) 18 mg/kg + Apatinib 150 mg/kg, d): Paclitaxel (albumin binding type) 16 mg/kg + Apatinib 150 mg/kg. The tumor growth curve was analyzed during the test. The apoptosis related protein (Bax, bcl-2), angiogenesis related protein (CD31, p-VEGFR-2) and invasion related protein(MMP-2) were analyzed by Western blot, Immunofluescence and Immunohistochemistry to analysis the effect of antitumor and side reactions Result (1) The IC-50 value of SKOV-3/DDP to paclitaxel (albumin binding type) was 45.53 ± 4.06 µmol/l, while the role of Apatinb was 50.66 ± 4.96umol/L(48 h). (2) the expressions of bcl-2(nab-P group, AP group), p-VEGFR-2(AP group), MMP-2(nab-P group, AP group) were higher than Control group, while Bax(nab-P group, AP group) lower (P < 0.01). (3) The invasion and migration of the cells decreased after the nab-P and AP treatment(P < 0.01). (4) nab-P combined with AP can increased their antitumor effect, according to Compusyn software, CL < 1. (5) After combined with AP, when nab-P were reduced dose in proper quantity, there were no obvious different in drug effect. (6) Reduced nab-P can increased nude mice’s quality of life. Conclusion Paclitaxel (albumin binding type), a chemotherapeutic agent, can play an anti-tumor role in drug-resistant ovarian cancer, but it can also reduce the tumor load and increase the expression of tumor vascular endothelial growth factor. When combined with Apatinib, the target drug of anti vascular endothelial growth factor, the two drugs have synergistic effect, which can improve the effect of single drug. In addition, when combined with Apatinib, the dosage of Paclitaxel (albumin binding type) can be appropriately reduced under the standard of recommended dosage to reduce the toxicity of chemotherapy drugs, without affecting the anti-tumor effect.

scholarly journals Nobiletin enhances Pirarubicin chemosensitivity of breast cancer cell MDA-MB-231 by regulating PER2

2021 ◽  
Vol 233 ◽  
pp. 02010
Author(s):  
Fahu Yuan ◽  
Xinghua Liao ◽  
Tongcun Zhang
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Scratch Test ◽  
Inhibitory Effect ◽  
Control Group ◽  
Potential Mechanism ◽  
Related Protein ◽  
Repair Ability ◽  
Apoptosis Related Protein ◽  
Related Proteins

To explore the potential mechanism of nobiletin in improving the sensitivity of breast cancer MDA-MB-231 cells to Pirarubicin. MDA-MB-231 cells were randomly divided into Control group, Nobiletin group, Pirarubicin group and Nobiletin co-Pirarubicin group. CCK-8 assay was used to detect the inhibitory effect of Nobiletin co-Pirarubicin on the proliferation of MDA-MB-231 cells, and the cell cycle and apoptosis of MDA-MB-231 cells were detected by flow cytometry. The effect of nobiletin combined with Pirarubicin on the migration and repair ability of MDA-MB-231 cells was detected by scratch test. Western blotting was used to determine the effect of nobiletin co-Pirarubicin on Period2 (PER2), apoptosis-related proteins Bax and BCL-2 expression levels in MDA-MB-231 cells. The results showed that the sensitivity of MDA-MB-231 cells to Pirarubicin was enhanced by nobiletin. Compared with Pirarubicin or nobiletin alone, Pirarubicin combined with nobiletin promoted the apoptosis of MDA-MB-231 cells and the expression of apoptosis-related protein Bax, and reduced the expression of apoptosis-inhibiting protein BCL-2. The combination of nobiletin and Pirarubicin significantly inhibited the proliferation and promoted the apoptosis of MDA-MB-231 cells, which may be mediated by the enhancement of PER2 expression by nobiletin.

scholarly journals OR-031 Mechanism of free-wheeling exercise attenuating muscle atrophy in SAMP8 mice

2018 ◽  
Vol 1 (2) ◽  
Author(s):  
Ying Zhang ◽  
Xianjuan Kou
Keyword(s):  
Skeletal Muscle ◽  
Protein Expression ◽  
Western Blot ◽  
Muscle Atrophy ◽  
Control Group ◽  
Related Protein ◽  
Autophagy Inhibitor ◽  
Apoptosis Related Protein ◽  
Related Proteins ◽  
Samp8 Mice

Objective Muscle atrophy is a decrease in the volume and number of skeletal muscle cells, decreased muscle strength, and increased connective tissue and fat as the patient ages, and clinical symptoms syndrome characterized by decreased physical function, falls, weakness, and varying degrees of disability. It is estimated that approximately 5-13% of the elderly over the age of 60 are affected by muscle atrophy. Studies have shown that increased myocyte apoptosis and decreased levels of autophagy are involved in the development of muscle atrophy. At present, appropriate exercise has been considered as an economical and convenient way to reduce muscle atrophy, delay aging, and activate autophagy. The molecular mechanism remains to be further studied. The aim of this study was to investigate the effect and mechanism of 8-week free-wheeling exercise on muscle atrophy in rapid aging model SAMP8 mice, in order to provide a theoretical basis for exercise training to improve muscle atrophy and inhibit apoptosis. Methods Twenty-four SAMP8 male mice of 7-month-old SPF were randomly divided into control group (Con), exercise group (V), and exercise combined with autophagy inhibitor chloroquine group (VQ), with 8 rats in each group. The Con group was fed routinely without any intervention for a total of 8 weeks. After 8 weeks, the mice were sacrificed by cervical dislocation, and the gastrocnemius muscle tissue was taken immediately. The apoptosis of skeletal muscle and the expression of aging-related proteins were detected by Western-blot. Test indicators include BAX, Ac-p53, p21, and p16. All experimental data were analyzed by statistical software and showed significant differences at P < 0.05. Results Western-blot results showed that compared with the Con group, the expression of apoptosis-related protein BAX and aging-related proteins AC-P53, p16, and p21 were significantly decreased in the skeletal muscle of the V group (P<0.01). In the VQ group, the protein expression of BAX was significantly increased (P<0.01), and the protein expression of AC-P53, p16 and p21 also increased (P<0.01). Conclusions The above Western-blot results indicate that free-wheeling exercise significantly inhibits skeletal muscle cell apoptosis in aged mice and reduces muscle atrophy.At the same time, under the action of free-wheeling exercise combined with autophagy inhibitor chloroquine, the apoptosis-related protein and aging-related protein increased abnormally, suggesting that free-running movement may inhibit apoptosis and delay muscle atrophy by activating autophagy. Further research is needed.

scholarly journals Combination of ω-3 Polyunsaturated Fatty Acids with 5-FU Inhibits SGC7901 Gastric Cancer Cell Growth via Rho-ROCK1 Signaling Pathway in Nude Mice

2021 ◽  
Author(s):  
Jiang Yiyan ◽  
Wang Keke ◽  
Lou Zhefeng ◽  
Hong Dan ◽  
Min Tao
Keyword(s):  
Gastric Cancer ◽  
Western Blot ◽  
Cancer Cells ◽  
Nude Mice ◽  
Protein Level ◽  
Combination Group ◽  
Control Group ◽  
Mrna Levels ◽  
Caspase 9 ◽  
Gastric Cancer Cells

Abstract Background: Gastric cancer is one of the most common malignancy with high mortality rate in the world. Systemic chemotherapy is thought to be an important treatment. However, due to the unsatisfactory efficiency and obvious side effects, it is urgent to detect new therapy strategy for gastric cancer. This study was aimed to investigate the effects and mechanisms of ω-3 polyunsaturated acids (PUFAs) combined with 5-FU on the growth of gastric cancer cells in nude mice. Methods: BALB/C nude mice were injected subcutaneously with SGC7901 gastric cancer cells to establish a tumor-bearing mouse model. The tumor growth in vivo was observed. Morphological of tumor specimens was observed by HE staining. The mRNA levels of RhoA, RhoC and ROCK1 in tumor tissues were detected by qPCR, and their protein levels were detected by immunofluorescence and Western Blot. Meanwhile, apoptosis –related proteins were also determined by Western Blot.Results: Compared with the NC control group, the tumor volume and weight in ω-3 PUFAs and 5-Fu groups were insignificantly lower, but significantly lower in the combination group. Compared with the abundant blood supply in the NC group, HE staining showed multifocal tumor necrosis in the three intervention groups, and this change was the most prominent in the combination group. And qPCR results showed that the mRNA levels of RhoA in the combination groups were significantly lower than this in the other groups. Immunofluorescence showed that the level of RhoA protein in the three intervention groups decline in varying degrees, especially in the combination group. Western Blot showed that the protein level of RhoA in the three intervention groups were significantly lower than those in the NC control group, especially in the combination group. Meanwhile, the protein level of ROCK1 in both 5-FU group and the combination group were significantly lower, especially in the combination group. Compared with the control group, the levels of Bcl-2 and Caspase-9 decreased in the combination group, the level of cleaved Parp was increased at the same time.Conclusion: ω-3 PUFAs combined with 5-FU may inhibit tumor growth through the Rho/ROCK pathway and promote apoptosis by down-regulating the levels of Bcl-2 and Caspase-9 and induce the increase of cleaved Parp level.

Lidocaine Inhibits Ovarian Cancer Cell Proliferation and Induces Apoptosis: An In Vitro Study

2020 ◽  
Vol 10 (5) ◽  
pp. 724-729
Author(s):  
Yaping Xu ◽  
Xiaoqin Fang ◽  
Xianjiang Wei
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Protein Expression ◽  
Western Blot ◽  
Cancer Cell ◽  
Blot Analysis ◽  
Group Treatment ◽  
Western Blot Analysis ◽  
Ovarian Cancer Cell ◽  
Control Group

Objective: The present study aimed to explore the effects and related mechanism of lidocaine on human ovarian cancer cell lines. Methods: Human ovarian cancer cell lines (SKOV3 and ES-2) were treated with different concentrations of lidocaine for different time. We treated SKOV3 and ES-2 cells using lidocaine then used MTT assay and flow cytometry to detect the cell proliferation and cell apoptosis. In addition, we used western blot analysis to explore the protein expression of Bax and Bcl-2 in SKOV3 and ES-2 cells. Western blot analysis and qRT-PCR were performed for the detection of EMT markers (E-cadherin, N-cadherin). The protein expression levels of TRAF3 and p-p65 in SKOV3 and ES-2 cells were determined by Western blot analysis. Results: Compared to the control group, 0.5, 1, 5, and 10 mM of lidocaine significantly inhibited ovarian cancer cell proliferation at different time points, while 0.1 mM of lidocaine had no significant effect. 1, 5 mM of lidocaine induced the cell apoptosis, and observably reduced expression of Bcl-2 protein, but improved Bax expression markedly compared with the control group. Treatment of lidocaine increased E-cadherin expression, but decreased N-cadherin expression when compared with control group. Treatment of lidocaine increased TRAF3 protein expression, but decreased p-p65 protein expression in ES-2 and SKOV3 cells. Conclusion: We demonstrated that lidocaine inhibited cell proliferation, induced apoptosis, and inhibited EMT in ovarian cancer cells via regulating TRAF3/NF-κB pathway.

Effect of Plant Polyphenol Genistein on Growth of Human Ovarian Carcinoma Transplanted Subcutaneously in Nude Mice

2013 ◽  
Vol 781-784 ◽  
pp. 587-590
Author(s):  
Bin Liu ◽  
Jie Yun Sun ◽  
Li Yu
Keyword(s):  
Ovarian Cancer ◽  
Ovarian Carcinoma ◽  
Nude Mice ◽  
Inhibitory Effect ◽  
Combination Group ◽  
Control Group ◽  
Weight Changes ◽  
Plant Polyphenol ◽  
Cancer Tumor ◽  
Human Ovarian Carcinoma

In this study we investigated the effect of plant polyphenol PTK inhibitor genistein on subcutaneous human transplant ovarian cancer tumor of nude mice. All 30 cases of nude mice are used to establish subcutaneous xenograft models of human ovarian cancer, and divided into 4 groups: Control group (containing 0.04% DMSO of saline); Genistein group (containing 0.2 mg / kg and 0.4 mg/kg two concentrations, subcutaneous injection); Cisplatin group (4 mg/kg, i.v.); Genistein and Cisplatin combination group. The transplanted tumor growth and weight changes of nude mice in different groups on 7, 14, 21 and 28 days were observed and histopathological examined. The results showed that the growth of SKOV3 xenograft tumor was significantly inhibited in 0.4 mg/kg Genistein group. Compared with control group, the tumor weights were decreased, the tumor volumes were reduced, and there was a significant increase in the area of necrosis, but no significant effects were showed on the weights of nude mice. 0.4 mg/kg Genistein (s.c.) in combination with 4 mg/kg Cisplatin (i.v.) enhanced the inhibitory effect. The results provide evidence for the potential usefulness of Genistein in the prevention and treatment of human ovarian carcinoma.

scholarly journals Sclerostin/Receptor Related Protein 4 and Ginkgo Biloba Extract Alleviates β-Glycerophosphate-Induced Vascular Smooth Muscle Cell Calcification By Inhibiting Wnt/β-Catenin Pathway

2019 ◽  
Vol 47 (Suppl. 1) ◽  
pp. 17-23 ◽  
Author(s):  
Jian Wang ◽  
Xiaobo Qiu ◽  
Tianhua Xu ◽  
Zitong Sheng ◽  
Li Yao
Keyword(s):  
Smooth Muscle ◽  
Western Blot ◽  
Ginkgo Biloba ◽  
Real Time Pcr ◽  
Negative Control Group ◽  
Ginkgo Biloba Extract ◽  
Negative Control ◽  
Control Group ◽  
Related Protein ◽  
Alizarin Red

Background: Abnormal mineral metabolism in patients with chronic kidney disease (CKD) may lead to vascular calcification, which is markedly associated with adverse events, including ischemic cardiac diseases and all-cause cardiovascular mortality. Thus, preventing and treating vascular calcification play an important role in improving the prognosis of CKD patients. Objectives: To investigate the potential functions of sclerostin and low-density lipoprotein receptor-related protein 4 (Lrp4) in alleviating the β-glycerophosphate (β-GP)-induced vascular smooth muscle cell (VSMC) calcification, and the protective effect of Ginkgo biloba extract (GBE). Methods: VSMC were extracted from Sprague-Dawley rat aorta and cultured in medium. The VSMCs were divided into 3 groups: (1) Negative control group, (2) β-GP group, in which the VSMCs were treated with β-GP, and (3) GBE and β-GP group, where the VSMCs were treated with both β-GP and GBE. The calcium nodules within the cells were examined by using Alizarin red S staining. The mRNA expression levels of β-catenin and bone gamma-carboxyglutamic-acid-containing proteins (BGP) were detected by real-time PCR. The protein levels of sclerostin and Lrp4 were determined by Western blot. Results: Alizarin red S staining showed that the VSMCs in β-GP group had a distinct orange-red precipitate when compared with VSMCs in the negative control group, while the orange-red precipitate of the GBE and β-GP group was significantly reduced compared to the β-GP group. Real-time PCR showed that the mRNA levels of β-catenin and BGP in VSMCs of β-GP group were significantly higher than those of the negative control group (p < 0.05); while they were significantly reduced in VSMCs of the GBE and β-GP group (p < 0.05). Western blot results showed that the expression of sclerostin in the β-GP group was significantly higher than that in the control group (p < 0.05), whereas Lrp4 was significantly lower than in control group (p < 0.05). Sclerostin in GBE and β-GP group was significantly reduced (p < 0.05), but Lrp4 was significantly elevated when compared with that of the β-GP group (p < 0.05). Conclusion: β-GP induced VSMC calcification by activating the Wnt/β-catenin signaling pathway. Sclerostin and Lrp4 were involved in β-GP-induced VSMC calcification and play an important role. GBE could alleviate VSMC calcification induced by β-GP through inhibiting the Wnt/β-catenin signaling pathway.

Cancer-IgG upregulation of radioresistance in non-small cell lung cancer patients and cell lines.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e21013-e21013
Author(s):  
Guangying Zhu ◽  
Xiongtao Yang ◽  
Wang Guohui ◽  
Jing You
Keyword(s):  
Western Blot ◽  
Tumor Cells ◽  
Clinical Stage ◽  
Scratch Test ◽  
Gene Expression Omnibus ◽  
The Cancer Genome Atlas ◽  
Normal Lung ◽  
Migration And Invasion ◽  
Control Group ◽  
Expression Levels

e21013 Background: Immunoglobulin G (IgG) is produced by B lymphocytes which is an important effector molecule of human immunity. However,our collaborators in China Peking University have found that malignant tumor cells can also express IgG (cancer-IgG). This study was designed to investigate the relationship between cancer-IgG and radiosensitivity and to explore its possible mechanisms in NSCLC. Methods: First, the expression of IGHG1 (immunoglobulin heavy constant gamma 1, IGHG1) and cancer-IgG in NSCLC and their relationship with clinicopathological features and prognosis were analyzed by TCGA(The Cancer Genome Atlas, TCGA), GEO(Gene Expression Omnibus, GEO) databases and immunohistochemistry. Secondly, after silencing the expression of cancer-IgG, cell scratch test, transwell, clone formation and apoptosis experiments were performed to observe the changes of EMT and radiation sensitivity of tumor cells after irradiation. Finally, the key proteins of PI3K/Akt signaling pathway and EMT-related proteins were detected by western blot and real-time PCR. Results: 1. The expression levels of IGHG1 and cancer-IgG in NSCLC were significantly higher than those in normal lung tissues, and high expression of IGHG1 and cancer-IgG were factors of poor prognosis. And cancer-IgG was closely related to clinical stage (P = 0.042), T stage (P = 0.044) and metastasis (P = 0.007). 2. Cell scratches and transwell experiments showed that the cell migration and invasion ability of si-control+IR group were significantly enhanced, but in si-Cancer-IgG+IR group were significantly lower than si-control+IR group. 3. Cloning formation experiments showed that the number of cell clones formed in the si-cancer-IgG group was significantly less than that in the control group under 2-6 Gy irradiation. Apoptosis results showed that the apoptosis rates of si-control group, si-Cancer-IgG group, si-control+IR group and si-Cancer-IgG group were 6.36±1.69%, 21.33±1.21%, 23.31±0.96% and 52.69±2.33% respectively. The apoptotic cells in the Si-cancer-IgG group were significantly more than the si-control group. (P < 0.01). 4. Western blot results showed that p-AKT, p-GSK3β, bcl2 and Vimentin were significantly decreased in si-Cancer-IgG+IR group compared with si-control+IR, and the expression levels of BAD and E-cadherin were increased. Conclusions: Cancer-IgG may mediate radiation-induced EMT and radiotherapy resistance through the PI3K/AKT pathway.

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