Objective To investigate the effect and underlying mechanism of Caudatin combined with Gefitinib on Gefitinib resistance induced by HGF in PC-9.Methods Model of EGFR-TKIs resistance in PC-9 cells was induced by exogenous HGF and co-cultured with MRC-5.Caudatin was tested as a drug resistant modulator to reverse the resistance of Gefitinib in PC-9 cells induced by HGF by MTT assay.Western blot was performed to observe the mechanism of Caudatin combined with Gefitinib reversing the resistance of PC-9 induced by HGF.Results The resistance of gefitinib to PC-9 was induced by exogenous HGF and co-cultured with MRC-5 which could reduce relative inhibitory rate ( P<0.05 ) .Neither caudatin ( 0-32 μM ) or Gefitinib (1μM) alone could significantly inhibit proliferation of PC-9 in the presence of HGF, which could be inhibited in a dose-dependent manner by Caudatin combined with Gefitinib ( P<0.05 ); Caudatin combined with Gefitinib down-regulated the phosphorylation levels of Met and PI3K/Akt simultaneously (P<0.05).Conclusion Caudatin could reverse the drug resistance of Gefitinib in PC-9 induced by HGF, the mechanism of which may be related to the inhibition of Met/PI3K/AKT pathway.

Glucose-6-phosphate dehydrogenase (Glucose-6-phosphate dehydrogenase, G6PD) is the rate-limiting enzyme of pentose phosphate pathway (PPP), which mainly maintains the balance of NADPH and intracellular redox reaction. Reducing G6PD activity or PPP dysfunction can prevent normal cell proliferation, and severe lack of G6PD can damage embryonic development and delay organ growth. At present, many studies have proved that abnormal activation of G6PD can lead to the enhancement of cell proliferation and adaptability of various types of cancer, and it is easy to cause drug resistance and increase the difficulty of clinical treatment. It has become an urgent need for clinical treatment to study the mechanism of G6PD in cancer cells and identify new potential drug therapeutic targets.

AngiotensinⅡ( AngⅡ) is the main functional bioac-tive peptide of the renin angiotensin system,and its basic function is to regulate salt-water homeostasis and blood pressure. Howev-er,recent studies have shown that AngⅡ plays a very important role in tumor formation and angiogenesis by acting on its type 1 receptor (AT1R) as a kind of potential growth factor. This arti-cle is a brief overview of the research on effects of AngⅡ-AT1R system in the process of tumor angiogenesis in recent years.

Lung is the breathing organ of the human body. The respiration of the lung enables the body to exchange oxygen with the outside world to maintain the body's life activities. The physiological properties of the lungs expose the mucosal surfaces of the lungs to harmful external irritants, including pathogens, allergens, harmful gases and smoke particles, which can lead to lung injury and infection. Induced by lung inflammation, immune cells (B lymphocytes and T lymphocytes) in the lungs gather around the bronchi, forming induced bronchial associated lymphoid tissue (iBALT). Studies have found that iBALT is involved in the pathological development of asthma, COPD, lung cancer and other lung diseases. iBALT is regulated by a variety of cytokines, but the formation process and its effect on disease remain unclear. The purpose of this paper is to review the formation factors of iBALT and its pathological status in lung, and to provide new treatment ideas for chronic pulmonary inflammatory diseases.

AIM: To investigate the effect of Shenqi fuzheng injection (SFI) on tumor immunity and its preliminary molecular mechanism. METHODS: The animal model of low glucose tumor microenvironment was established by B16-PKM2-OE; the level of interleukin-2(IL-2) and interferon-γ(IFN-γ), CD40L and transforming growth factor-β1(TGF-β1) were detected by ELISA kit; the expressions of glucose transporter-1 (Glut-1) and key enzymes of glycolysis ( HK, PFK and PK ) in CD4

AIM: To analyze the clinical characteristics of anticoagulant rat poisoning and vitamin K

AIM: To investigate the function and mechanism of quercetin (Que) in anti-fibrosis in vitro and in vivo from the perspective of interfering with the glycolysis of renal interstitial fibroblasts. METHODS: ln vivo experiments, mice were administered in groups, kidneys were dissected, weighed and examined histopathologically and biochemically; ln vitro experiments, rat normal renal fibroblasts (NRK-49F cells) were treated with different reagents, proteins were extracted, and NRK-49F cell activation indicators such as α-smooth muscle actin (α-SMA) were detected by protein immunoblotting (Western Blot). The expression of the proteins, such as proliferating cell nuclear antigen (PCNA), was examined by protein immunoblotting (Western Blot), and the effect of Que on glucose uptake in NRK-49F cells induced by transforming growth factor-β (TGF-β1) and epidermal growth factor (EGF) was examined by fluorescence assay; the lactate content of cells in different experimental groups was examined by lactate assay kit; the effect of Que on glucose uptake in NRK-49F cells induced by TGF-β1 and EGF was examined by fluorescence quantitative PCR. EGF-induced mRNA of hexokinase (HK2), phosphofruc-tokinase 1 (PFK1) and muscle pyruvate kinase isozyme 2 (PKM2), key enzymes of glycolysis in NRK-49F cells. RESULTS: Compared with the UUO group, the morphological structures of kidney tissues in the Que administration group were all alleviated to different degrees, which were related to the inhibition of glycolysis, and the serum levels of urea nitrogen (BUN) and blood creatinine (Scr) in mice showed a significant downward trend; lactate production and glucose uptake in NRK-49F cells were gradually reduced, and Que affected TGF-β1 and EGF-induced RIF of mRNA levels of key enzymes of glycolysis gradually decreased and were associated with PKM2. CONCLUSION: Que inhibits PKM2 enzyme activity and glycolysis in NRK-49F cells and reduces TGF-β1-induced myofibroblast activation.

OBJECTIVE: To observe the cell death pattern induced by gefitinib in non-small cell lung cancer A549 and H1975 cells and explore the possible mechanism in light of glycolysis. METHODS: The inhibitory effects of gefitinib at 20, 30, or 40 μmol/L in A549 cells and at 20, 40, or 80 μmol/L in H1975 cells were examined using MTT assay. The changes of lactic acid level in the cells were determined with a lactic acid kit, and the expression levels of glycolysis-related proteins (PKM2 and HK2) and the proteins in PI3K-Akt-mTOR signaling pathway were detected using Western blotting. 2-NBDG was used for detecting glucose uptake capacity of the cells, and ATP kit was used to detect the intracellular ATP level. The mitochondrial membrane potential of the cells was examined with the JC-1 kit, and cell apoptosis was analyzed with Annexin V-FITC/PI double staining. The relative expression levels of the apoptotic proteins Bax and Bcl-2 and the autophagy marker protein LC3B were detected with Western blotting. RESULTS: MTT assay showed that gefitinib inhibited the proliferation of A549 and H1975 cells in a time- and dose-dependent manner ( < 0.05). The IC of gefitinib at 24, 48 and 72 h was 48.6, 28.6 and 19.7 μmol/L in A549 cells and was 321.6, 49.1 and 14.6 μmol/L in H1975 cells, respectively. Gefitinib significantly lowered intracellular lactic acid level of the cells ( < 0.05) and down-regulated the expressions of PKM2 and HK2 proteins ( < 0.05) and PI3K-Akt-mTOR signaling pathway-associated proteins ( < 0.05). Gefitinib obviously inhibited glucose uptake and ATP levels in both A549 and H1975 cells ( < 0.05). Treatment with gefitinib induced obviously enhanced apoptosis in the cells, resulting in apoptosis rates of (10.77± 1.0)%, (14.5±0.4)%, (17.4±0.2)% and (32.1±0.6)% at 0, 20, 30 and 40 μmol/L in A549 cells ( < 0.05) and of (10.5±0.6)%, (13.2± 0.92)%, (18.9±0.98)% and (35.1±1.4)% at 0, 20, 40 and 80 μmol/L in H1975 cells, respectively ( < 0.05). The protein expression of Bax increased and that of Bcl-2 decreased following gefitinib treatment in the cells ( < 0.05). Gefitinib significantly increased autophagy in A549 and H1975 cells as shown by increased LC3B expressions following the treatment ( < 0.05). CONCLUSIONS Gefitinib can inhibit the proliferation, induce apoptosis and increase autophagy in A549 and H1975 cells. Gefitinib induces apoptosis of the cells possibly by affecting glycolysis and PI3K-Akt-mTOR signaling pathway.
Apoptosis ; Carcinoma, Non-Small-Cell Lung ; Cell Line, Tumor ; Cell Proliferation ; Gefitinib ; Glycolysis ; Humans ; Lung Neoplasms ; Phosphatidylinositol 3-Kinases

OBJECTIVES: Lung cancer is one of the most common malignant tumors in the world, and its lethality ranks the first among many malignant tumors. For non-small cell lung cancer (NSCLC) patients, due to the high mortality rate, the overall 5-year survival rate is less than 15%. When NSCLC undergoes local invasion, the 5-year survival rate is only 20%, and it is even lower when distant metastasis occurs up to 4%. Almonertinib is an innovative drug independently researched and developed by China with independent intellectual property rights. As an epidermal growth factor receptor tyrosine kinase inhibitor, almonertinib is mainly used for locally advanced or metastatic NSCLC patients with epidermal growth factor receptor (EGFR) T790M mutation. This study aims to investigate the effects of almonertinib on the proliferation, invasion and migration of NSCLC cells in vitro. METHODS: NSCLC cells H1975 and PC-9 were cultured in vitro. The effects of almonertinib on the proliferation, apoptosis, invasion, and migration of H1975 and PC-9 cells were detected by CCK-8 assay, apoptotic assay and Transwell assay. The expression of invasion and migration related proteins was detected by Western blotting. RESULTS: The CCK-8 experiment showed that almonertinib inhibited the proliferation of H1975 and PC-9 cells in a time- and dose-dependent manner. The IC CONCLUSIONS Almonertinib can inhibit the proliferation, invasion, and migration of NSCLCH1975 and PC-9 cells in vitro and vivo, and promote the apoptosis of H1975 and PC-9 cells. The underlying mechanism may be related to the inhibition of tumor cell epithelial mesenchymal transformation and metalloproteinase expression.
Acrylamides ; Animals ; Apoptosis ; Carcinoma, Non-Small-Cell Lung/drug therapy* ; Cell Line, Tumor ; Cell Proliferation ; Drug Resistance, Neoplasm ; ErbB Receptors/genetics* ; Humans ; Indoles ; Lung Neoplasms ; Mice ; Mice, Nude ; Mutation ; Protein Kinase Inhibitors/pharmacology* ; Pyrimidines

OBJECTIVE: To investigate the inhibitory effect of epidermal growth factor receptor tyrosine kinase inhibitor (EGFRTKI) HS-10296 on the proliferation of triple-negative breast cancer (TNBC) MDA-MB-231 cells and explore the possible molecular mechanism. METHODS: MDA-MB-231 cells were treated with HS-10296 for 24, 48, or 72 h, and CCK-8 assay was used to assess the changes in the cell viability. The inhibitory effect of HS-10296 on cell proliferation was determined by clonogenic assay. JC-1 and flow cytometry were employed for analyzing the cell apoptosis, and the ultrastructure of the cells was observed under electron microscope. After pretreatment with autophagy inhibitor chloroquine (CQ), MDA-MB-231 cells were divided into control group, CQ treatment group, HS-10296 (4 and 6 μmol/L) treatment groups and combined treatment groups, and the sensitivity of the treated cells to HS-10296 was determined using CCK-8 assay. The effects of HS-10296 on EGFR pathway and apoptosis- and autophagy-related proteins in MDA-MB-231 cells were investigated using Western blotting. RESULTS: HS-10296 significantly inhibited the proliferation of MDA-MB-231 cells with IC values at 24, 48 and 72 h of 8.393, 2.777 and 2.016 μmol/L, respectively. JC-1 and flow cytometry showed that HS-10296 induced obvious apoptosis of MDA-MB-231 cells, which showed an apoptosis rate of (21.63 ± 2.97)% following treatment with 8 μmol/L HS-10296. Autophagy vesicles were observed in the cells treated with HS-10296 under electron microscope. In MDA-MB-231 cells pretreated with CQ, inhibition of autophagy significantly enhanced HS-10296-induced cell death. Western blotting showed that the apoptosis-related protein caspase-3 was activated after HS-10296 treatment to cut its substrate PARP. The expression of autophagy-related protein light chain 3B (LC3B) was significantly enhanced after HS-10296 treatment ( < 0.01), which also resulted in inhibited phosphorylation of EGFR and AKT proteins in the cells. CONCLUSIONS HS-10296 can inhibit the proliferation and induce autophagy and apoptosis of MDA-MB-231 cells by inhibiting the EGFR/PI3K/AKT signaling pathway.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Autophagy ; drug effects ; Breast Neoplasms ; drug therapy ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; ErbB Receptors ; metabolism ; Humans ; Protein Kinase Inhibitors ; pharmacology ; Signal Transduction ; drug effects