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Aim To explore the effeet of the extract of Celastrus orbiculatus extract on the proliferation anrl ap- optosis of multiple myeloma eells (U-1996) and its molecular mechanism.Methods The U-1996 eells were divided into normal control ( NC ) group, the southern snake vine extraet group, si-NC group, si- LINC00472 group, pcDNA-NC group, pcDNA- LINC00472 group, southern snake vine extraet + si-NC group,and southern snake vine extraet + si-LINC00472 group.Heal-time quantitative PCR was used to deteet LINC00472 expression, CCK-8 and flow cytometry to deteet eell proliferation and apoptosis, Western blot to deteet the expression levels of phosphorvlated phos- phatidvlinositol-3 -hydroxykinase (p-PI3K) and phos- phorylated protein kinase B ( p-Akt).Results After treated with Celastrus orbieulatus extract, the viability of U-1996 eells and the expression of p-PI3K and p- Akt were significantly reduced, the apoptotie rate and the expression of LINC0047 were significantly raised ( P < 0.05 ).After inhibiting the expression of LINC00472, the viability of U-1996 eells and the ex¬pression of p-PBK and p-Akt significantly inereased, the apoptotie rate and the expression of LINC0047 sig¬nifieantly deereased ( P <0.05 ).After overexpression of LINC00472, the viability of U-1996 eells and the ex¬pression of p-PI3K and p-Akt were signifieantly re- dueed, the apoptotie rate and the expression of LINC0047 significantly inereased ( P <0.05 ).Inhibi¬ting the expression of LINC0047 can reverse the effects of Celastrus orbiculatus extract on proliferation , apopto- sis and PI3K/AKT signaling pathway of U-1996 cells (P < 0.05 ).Conclusions Celastrus orbiculatus ex¬tract can inhibit the proliferation of multiple myeloma cells and induce apoptosis through up-regulating LINC00472 to inhibit PI3K/Akt signaling pathway.
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ObjectiveTo study the inhibitory effect of Celastrus orbiculatus extract （COE） on gastric cancer cells， to clarify the specific mechanism of COE promoting the apoptosis of gastric cancer cells by affecting the mitochondrial structure and function， and to provide an experimental basis for the further development and clinical application of C. orbiculatus. MethodBrdu staining combined with flow cytometry and Annexin V-fluorescein isothiocyanate （AnnexinV-FITC） staining combined with flow cytometry were employed to detect the effects of COE （20， 40， 80 mg·L-1） on the proliferation and apoptosis of gastric cancer cells， respectively. The changes in mitochondrial membrane potential were detected with JC-1 mitochondrial membrane potential assay kit. The expression of apoptosis-associated proteins including B-cell lymphoma-2 （Bcl-2）， B-cell lymphoma-xL （Bcl-xL）， Bcl-2-associated X （Bax）， and cysteine aspartutespecific protease-3 （Caspase-3） in gastric cancer cells was determined by Western blot. Transmission electron microscopy was employed to detect changes in the mitochondrial microstructure of gastric cancer cells exposed to COE. Western blot was employed to measure the expression of mitochondrial marker proteins ［superoxide dismutase 1 （SOD1）， voltage-dependent anion channel （VDAC）， prohibitin 1 （PHB1）， and heat shock protein 60 （HSP60）］ in gastric cancer cells. ResultCompared with the control group， COE （40， 80 mg·L-1） inhibited the proliferation and promoted the apoptosis of gastric cancer cells （P<0.05）. Furthermore， COE reduced the mitochondrial membrane potential of gastric cancer cells. Compared with the control group， COE （20， 40， 80 mg·L-1） up-regulated the expression of Bax and Caspase-3 which promoted apoptosis of gastric cells （P<0.05， P<0.01）， and COE at 40 and 80 mg·L-1 down-regulated the expression of Bcl-2 and Bcl-xL which inhibited the apoptosis of gastric cancer cells （P<0.01）. The results of transmission electron microscopy showed that COE changed the microstructure of gastric cancer cells， which led to the appearance of vacuoles in the cell membrane and mitochondria and damaged the mitochondrial structure. Compared with the control group， COE （20， 40， 80 mg·L-1） changed the expression of mitochondrial marker proteins. Specifically， it up-regulated the expression of SOD1 involved in stress response （P<0.05， P<0.01） and down-regulated that of VDAC， PHB1， and HSP60 associated with mitochondrial stability and permeability （P<0.01）. ConclusionCOE can significantly inhibit the proliferation and promote the apoptosis of gastric cancer cells. It may activate the mitochondrial apoptosis pathway by destroying the mitochondrial structure and function of gastric cancer cells.
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ObjectiveTo study the inhibitory effect of Celastrus orbiculatus extract （COE） on gastric cancer cells， to clarify the specific mechanism of COE promoting the apoptosis of gastric cancer cells by affecting the mitochondrial structure and function， and to provide an experimental basis for the further development and clinical application of C. orbiculatus. MethodBrdu staining combined with flow cytometry and Annexin V-fluorescein isothiocyanate （AnnexinV-FITC） staining combined with flow cytometry were employed to detect the effects of COE （20， 40， 80 mg·L-1） on the proliferation and apoptosis of gastric cancer cells， respectively. The changes in mitochondrial membrane potential were detected with JC-1 mitochondrial membrane potential assay kit. The expression of apoptosis-associated proteins including B-cell lymphoma-2 （Bcl-2）， B-cell lymphoma-xL （Bcl-xL）， Bcl-2-associated X （Bax）， and cysteine aspartutespecific protease-3 （Caspase-3） in gastric cancer cells was determined by Western blot. Transmission electron microscopy was employed to detect changes in the mitochondrial microstructure of gastric cancer cells exposed to COE. Western blot was employed to measure the expression of mitochondrial marker proteins ［superoxide dismutase 1 （SOD1）， voltage-dependent anion channel （VDAC）， prohibitin 1 （PHB1）， and heat shock protein 60 （HSP60）］ in gastric cancer cells. ResultCompared with the control group， COE （40， 80 mg·L-1） inhibited the proliferation and promoted the apoptosis of gastric cancer cells （P<0.05）. Furthermore， COE reduced the mitochondrial membrane potential of gastric cancer cells. Compared with the control group， COE （20， 40， 80 mg·L-1） up-regulated the expression of Bax and Caspase-3 which promoted apoptosis of gastric cells （P<0.05， P<0.01）， and COE at 40 and 80 mg·L-1 down-regulated the expression of Bcl-2 and Bcl-xL which inhibited the apoptosis of gastric cancer cells （P<0.01）. The results of transmission electron microscopy showed that COE changed the microstructure of gastric cancer cells， which led to the appearance of vacuoles in the cell membrane and mitochondria and damaged the mitochondrial structure. Compared with the control group， COE （20， 40， 80 mg·L-1） changed the expression of mitochondrial marker proteins. Specifically， it up-regulated the expression of SOD1 involved in stress response （P<0.05， P<0.01） and down-regulated that of VDAC， PHB1， and HSP60 associated with mitochondrial stability and permeability （P<0.01）. ConclusionCOE can significantly inhibit the proliferation and promote the apoptosis of gastric cancer cells. It may activate the mitochondrial apoptosis pathway by destroying the mitochondrial structure and function of gastric cancer cells.
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To explore the effects of Celastrus orbiculatus extract (COE) on the expressions of apoptosis-related proteins and growth of human gastric adenocarcinoma SGC-7901 cell xenografts in nude mice. Thirty-five mice with subcutaneous xenografts of a gastric cancer cell line (SGC-7901) were randomly divided into five groups: a negative control group, a positive control group (xeloda), and low-dose, medium-dose, and high-dose COE groups (10, 20, and 40 mg/kg, respectively). There were seven mice in each group. Tumor volume, tumor weight, and body weight were measured to draw up tumor growth curves and the tumor weight inhibition rates were calculated. The apoptosis rates were measured by TUNEL staining method and the expressions of p53, Bcl-2, and Bax proteins were detected by immunohistochemical staining and Western blotting assay. The COE could inhibit the growth of SGC-7901 xenografts in a dose-dependent manner. The tumor inhibitory rates were 33.3%, 46.8%, and 57.7% when treated with 10, 20, and 40 mg/kg COE, respectively. TUNEL assay showed that COE presented with apparently more apoptosis than the negative control group. The results of immunohistochemical staining showed the expressions of p53 and Bax proteins had a trend of up-regulation, while the expression of Bcl-2 protein had a trend of down-regulation in nude mice after treatment with COE. Compared with the negative control group, the expressions of p53 and Bax were significantly up-regulated, while the expression of Bcl-2 was down-regulated in the high-dose COE group. Western blotting analysis showed the similar results to immunohistochemical staining. COE could significantly inhibit the growth of SGC-7901 cell xenografts in nude mice, which might be related with the up-regulation of the expressions of p53 and Bax and down-regulation of the expression of Bcl-2.