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1.
Article in Chinese | WPRIM | ID: wpr-855913

ABSTRACT

AIM: To investigate the effects of silencing carbonic anhydrase 1 (CA1) on proliferation, apoptosis, invasion and migration of human lung cancer A549 cells. METHODS: CA1-specific siRNA (si-CA1 group) and negative control (si-NC group) were transfected into lung cancer A549 cells by lipofection. The A549 cells transfected with empty liposome were used as blank control group. Real-time quantitative PCR (qPCR) and Western blot (Western blot) were used to detect the expression of CA1 mRNA and protein. Cell counting kit method (CCK-8), flow cytometry and Transwell assay were used to detect proliferation and apoptosis of A549 cells, invasion and migration capabilities. RESULTS:qPCR and Western blot showed that the expression levels of CA1 mRNA and protein in A549 cells transfected with CA1 siRNA were significantly down-regulated (P0.05). CONCLUSION: Silencing CA1 can inhibit the proliferation, invasion and migration of lung cancer A549 cells and promote cell apoptosis.

2.
Article in Chinese | WPRIM | ID: wpr-846423

ABSTRACT

Objective: To prepare polydopamine-modified elemene-loaded mesoporous silica nanoparticles (D/MSN-ELE), and conduct research on formulation process optimization, quality evaluation, in vitro release, in vitro antitumor activity, and ability to promote apoptosis. Methods: Elemene-loaded mesoporous silica nanoparticles (MSN-ELE) were prepared by solution adsorption method, D/MSN-ELE and polydopamine-modified mesoporous silica nanoparticles (D/MSN) were prepared by polymerization. The morphology of the nanoparticles was characterized by transmission electron microscopy. The PDA graft ratio was calculated by thermogravimetric analysis. The loading and encapsulation efficiency of D/MSN-ELE were evaluated using HPLC, the dialysis bag method was used to investigate the release characteristics in vitro of D/MSN-ELE. MTT staining was used to analyze the cytotoxicity of different nanoparticles on HELF and A549 cells. Flow cytometry was used to detect the levels of D/MSN-ELE reactive oxygen species and mitochondrial membrane potential. Results: The optimal preparation process was the drug loading ratio of 6:1, the temperature was 50℃, and the time was 8 h. The D/MSN-ELE prepare under the process condition have a were uniform distribution with a particle size of (288.70 ± 3.88) nm. The average drug loading and encapsulation efficiency were (11.58 ± 0.73)% and (59.82 ± 0.57)%, respectively. In vitro drug release was pH-responsive, and cumulative drug release increased with decreasing pH. The half-lethal concentrations of ELE, MSN-ELE and D/MSN-ELE on A549 cells were 91.29, 27.56 and 6.02 μg/mL, respectively. The detection results of reactive oxygen species and mitochondrial membrane potential further indicated that drug-loaded nanoparticles were able to promote tumor target cell apoptosis. Conclusion: D/MSN-ELE under the optimized process has a higher drug loading, pH-responsive drug release and greatly enhanced antitumor activity. This study provides further experiments basis for tumor-targeted delivery of elemene drugs based on mesoporous silica nanoparticles.

3.
Chinese Pharmacological Bulletin ; (12): 181-186, 2019.
Article in Chinese | WPRIM | ID: wpr-857279

ABSTRACT

Aim: To study the induction of apoptotic effect of sodium selenite on human lung cancer A549 cells and its mechanisms. Methods: A549 cells were exposed to different concentrations of sodium selenite for 24 h. MTT assay was applied to determine A549 cell proliferation. Inverted fluorescence microscope was used to investigate the morphological changes in A549 cells. Flow cytometry analysis was applied to assess the apoptotic rates of A549 cells. Laser confocal microscope was employed to measure the reactive oxygen species (ROS) fluorescence intensity. A multi-detection reader was used to determine the antioxidant parameter. Western blot was utilized to detect the expression of Keapl, Nrf2, HO-1 and Nrf2 in cytoplasm and nucleus. Results: MTT results showed that sodium selenite inhibited the proliferation of A549 cells in a concentration-dependent manner. After treatment with sodium selenite for 24 h, the apoptotic rate of A549 cells was markedly increased through Hoechst 33342 staining and flow cytometry measurement. Sodium selenite significantly up-regulated ROS and malondialdehyde (MDA) content and down-regulated the levels of superoxide dismutase (SOD) and glutathione (GSH). Meanwhile, sodium selenite treatment also reduced the expressions of Keapl, Nrf2 and HO-1 at protein levels and inhibited Nrf2 protein nuclear translocation in A549 cells. Conclusions: Treatment with sodium selenite induces A549 cells apoptosis, which may contribute to the anti-proliferation activity, induction of apoptosis and regulation of oxidative stress reaction and Keapl/Nrf2/ARE antioxidative signaling pathway expression.

4.
China Pharmacy ; (12): 1346-1349, 2016.
Article in Chinese | WPRIM | ID: wpr-504405

ABSTRACT

OBJECTIVE:To study the inhibitory mechanism of timosaponin B-Ⅱ(TB-Ⅱ) on the proliferation and migration of human lung cancer A549 cells. METHODS:A549 cells were treated with TB-Ⅱ [0(blank control),1,10 and 100 μg/ml] for 48 h,and total RNA and total protein were extracted respectively. Real time fluorescence quantitative-PCR and Western blot were used to detect mRNA and protein levels of IL-18. IL-18 in A549 cells was silenced by transfection;the expression of IL-18 mRNA and protein were compared among untransfection group,negative control group and transfection group;and then human lung can-cer A549 cells with silenced gene were treated with 10 μg/ml TB-Ⅱ for 24,48 and 72 h. The activity of cell proliferation was de-tected with CCK-8,and the change of cell migration ability was observed by streak method. RESULTS:Compared with blank con-trol,the expression of IL-18 mRNA and protein in A549 cells all increased after treated with TB-Ⅱ(P<0.05 or P<0.01),and were positively correlated with concentration. Compared with untransfection group,the expression of IL-18 mRNA and protein de-creased in transfection group(P<0.01). Compared with untransfected cell treated with TB-Ⅱ,the viability and migration ability of A549 cells with transfection gene increased after treated with TB-Ⅱ for 72 h(P<0.01). CONCLUSIONS:TB-Ⅱ can inhibit the proliferation and migration of A549 cells by up-regulating IL-18 gene expression.

5.
China Pharmacy ; (12): 906-909, 2016.
Article in Chinese | WPRIM | ID: wpr-504326

ABSTRACT

OBJECTIVE:To study the effects of Thymalfasin for injection on the apoptosis of human lung cancer A549 cells. METHODS:After treated with 0(blank control),25,50,100,200 and 400 mg/L Thymalfasin for injection for 24,48 and 72 h, the cell proliferation inhibitory rate was analyzed with MTT and calculated. After treated with 0(blank control),50 and 100 mg/L Thymalfasin for injection for 48 h,cell apoptosis was detected by flow cytometry,and the expression of Caspase-3,Bcl-2 and Bax and the phosphorylation level of Akt were deteced by Western blot. RESULTS:Compared with blank control group,proliferation in-hibitory rate of A549 cells increased after treated with Thymalfasin for injection,in concentration and time-dependent manner(P<0.05). The apoptotic rate of A549 cells increased after treated with Thymalfasin for injection 50,100 mg/L for 48 h (P<0.05). The expression of Caspase-3 increased while the Bcl-2/Bax and phosphorylation level of Akt decreased in A549 cells after treated with Thymalfasin for injection 100 mg/L (P<0.05). CONCLUSIONS:Thymalfasin for injection can inhibit the proliferation of A549 cells by activating Caspase-3,decreasing Bcl-2/Bax ratio,inhibiting Akt signal pathway and induce the apoptosis of A549 cells.

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