Journal: Frontiers in Immunology

Article Title: A comprehensive landscape analysis of autophagy in cancer development and drug resistance

doi: 10.3389/fimmu.2024.1412781

Figure Lengend Snippet: Several potential anticancer drugs (dihydroartemisinin and artesunate) in reversing the doxorubicin resistance of breast cancer in vitro . (A) The effect of dihydroartemisinin, artesunate, and doxorubicin on the proliferation inhibition rate of MCF-7 parental and its doxorubicin-resistant cell, MCF-7/ADM, was quantified by the CCK-8 method. Inhibition rate was calculated as the difference in optical density (OD) at 450 nm between the control and experimental group (OD 450 ) divided by OD 450 in the control group and then multiplied by 100%. (B) RT-qPCR analysis of the levels in cells, and corresponding quantification of the ATG7 and LC3B mRNA levels. (C–E) An immunofluorescence assay was used to detect ABCG2, ATG7, and LC3B proteins in MCF-7/ADM cells. MCF-7/ADM was untreated or treated with ADM (2 μmol/L)/ART (20μmol/L)/DHA (20 μmol/L) and combined group (ART+ADM) for 48 h Proteins were stained green, and nuclei were stained blue with 4,6-diamidino-2-phenylindole (DAPI). The images were captured at 200× magnification. (F) The immunofluorescence results (using mean fluorescence intensity) to reflect the changes in protein expression levels. (G) Western blot analysis of the ATG7 protein levels in MCF-7/ADM, and corresponding quantification of the ATG7 protein. * p < 0.05, ** p < 0.01 vs. the control group.

Article Snippet: As a continuation of the previous research , the cells were treated with different concentrations of dihydroartemisinin (DHA, MedChemExpress, MCE, USA, CAS: 71939-50-9) or artesunate (ART, MCE, CAS: 88495-63-0) for 48 h, including 5, 10, 20, and 40 μmol/L (μM) ART; 10, 20, 40, and 80 μmol/L (μM) DHA; and different concentrations of DOX hydrochloride (ADM, MCE, CAS: 25316-40-9).

Techniques: In Vitro, Inhibition, CCK-8 Assay, Control, Quantitative RT-PCR, Immunofluorescence, Staining, Fluorescence, Expressing, Western Blot

Journal: Oncology Reports

Article Title: Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

doi: 10.3892/or.2023.8591

Figure Lengend Snippet: Cytotoxic activity of cisplatin, artesunate and iron on head and neck squamous cell carcinoma cells. (A and B) Cell viability was assessed at 72 h after exposure to different concentrations of (A) cisplatin (n=3), (B) artesunate (n=3) and (C) Fe(NO 3 ) 3 (n=3), followed by a WST-1 assay. All assays were performed in triplicate and the data are presented as the mean ± standard deviation. *P<0.0001, **P<0.005 and ***P<0.05 vs. control.

Article Snippet: The drugs used were artemisinin (CAS RN ® : 63968-64-9; cat. no. A2118; Tokyo Chemical Industry Co., Ltd.), deoxyartemisinin (CAS RN ® : 72826-63-2; cat. no. D232150; Toronto Research Chemicals), dihydroartemisinin (CAS RN ® : 81496-82-4, cat. no. D3793; Tokyo Chemical Industry Co., Ltd.), artesunate (CAS RN ® : 88495-63-0, cat. no. A2191; Tokyo Chemical Industry Co., Ltd.), and cisplatin (Randa Injection; Nippon Kayaku Co., Ltd.).

Techniques: Activity Assay, WST-1 Assay, Standard Deviation, Control

Journal: Oncology Reports

Article Title: Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

doi: 10.3892/or.2023.8591

Figure Lengend Snippet: Impact of combined treatment with ARTS and CDDP on cytotoxicity in HNSCC cells. (A) HNSCC cells (UM-SCC-81) were treated with various concentrations of CDDP combined with ARTS (1.56 or 3.125 µM) and Fe (0.02 mM) for 72 h followed by WST-1 assay. All assays were performed in triplicate. The line graph demonstrates the cytotoxic activity of the combined treatment. The cytotoxic activity of ARTS (1.56 or 3.125 µM) and Fe (0.02 mM) alone is also shown as points in the figure. *P<0.001 vs. cisplatin treatment at each concentration. (B) Cytotoxic activity of CDDP (0.25 µg/ml), ARTS (3.1 µM) and Fe (0.02 mM) on HNSCC cells (UM-SCC-23 and UM-SCC-81) was examined, alone and in combination. *P<0.01 vs. control. All assays were performed in triplicate and the data are presented as the mean ± standard deviation. ARTS, artesunate; CDDP, cisplatin; HNSCC, head and neck squamous cell carcinoma; Fe, Fe(NO 3 ) 3 .

Article Snippet: The drugs used were artemisinin (CAS RN ® : 63968-64-9; cat. no. A2118; Tokyo Chemical Industry Co., Ltd.), deoxyartemisinin (CAS RN ® : 72826-63-2; cat. no. D232150; Toronto Research Chemicals), dihydroartemisinin (CAS RN ® : 81496-82-4, cat. no. D3793; Tokyo Chemical Industry Co., Ltd.), artesunate (CAS RN ® : 88495-63-0, cat. no. A2191; Tokyo Chemical Industry Co., Ltd.), and cisplatin (Randa Injection; Nippon Kayaku Co., Ltd.).

Techniques: WST-1 Assay, Activity Assay, Concentration Assay, Control, Standard Deviation

Journal: Oncology Reports

Article Title: Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

doi: 10.3892/or.2023.8591

Figure Lengend Snippet: ARTS induces the accumulation of HNSCC cells in the S/G2 M phase, which was further enhanced by the combination of ARTS, Fe and CDDP. (A) HNSCC cells (UM-SCC-23) were treated with and without a combination of ARTS (3.1 µM), Fe (0.02 mM) and CDDP (0.3 µg/ml) for 72 h, and cell cycle distribution was examined by flow cytometry. The experiments were performed independently three times and the figure shows a representative experiment. (B) Data in the columns are presented as the mean ± standard deviation of three independent experiments. The results of statistical analysis are depicted for S/G2-M. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. ARTS, artesunate; HNSCC, head and neck squamous cell carcinoma; CDDP, cisplatin; Fe, Fe(NO 3 ) 3 .

Article Snippet: The drugs used were artemisinin (CAS RN ® : 63968-64-9; cat. no. A2118; Tokyo Chemical Industry Co., Ltd.), deoxyartemisinin (CAS RN ® : 72826-63-2; cat. no. D232150; Toronto Research Chemicals), dihydroartemisinin (CAS RN ® : 81496-82-4, cat. no. D3793; Tokyo Chemical Industry Co., Ltd.), artesunate (CAS RN ® : 88495-63-0, cat. no. A2191; Tokyo Chemical Industry Co., Ltd.), and cisplatin (Randa Injection; Nippon Kayaku Co., Ltd.).

Techniques: Flow Cytometry, Standard Deviation

Journal: Oncology Reports

Article Title: Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

doi: 10.3892/or.2023.8591

Figure Lengend Snippet: ARTS induces apoptosis in HNSCC cells, which was further enhanced by the combination of ARTS, Fe and CDDP. HNSCC cells (UM-SCC-23) were treated with or without a combination of ARTS (37.5 µM), CDDP (1.0 µg/ml) and Fe (0.01 mM) for 48 or 72 h, and apoptosis was examined by flow cytometry using annexin V and propidium iodide. (A) Flow cytometric profiles of live, apoptotic and necrotic cells at 48 and 72 h after treatment. (B) Data in the columns are presented as the mean ± standard deviation. *P<0.01 vs. control. ARTS, artesunate; HNSCC, head and neck squamous cell carcinoma; CDDP, cisplatin; Fe, Fe(NO 3 ) 3 ; FITC, fluorescein isothiocyanate.

Article Snippet: The drugs used were artemisinin (CAS RN ® : 63968-64-9; cat. no. A2118; Tokyo Chemical Industry Co., Ltd.), deoxyartemisinin (CAS RN ® : 72826-63-2; cat. no. D232150; Toronto Research Chemicals), dihydroartemisinin (CAS RN ® : 81496-82-4, cat. no. D3793; Tokyo Chemical Industry Co., Ltd.), artesunate (CAS RN ® : 88495-63-0, cat. no. A2191; Tokyo Chemical Industry Co., Ltd.), and cisplatin (Randa Injection; Nippon Kayaku Co., Ltd.).

Techniques: Flow Cytometry, Standard Deviation, Control

Journal: Oncology Reports

Article Title: Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

doi: 10.3892/or.2023.8591

Figure Lengend Snippet: ARTS reduces Rb and pRb levels in HNSCC cells. (A) Western blotting was performed for pRb and Rb. ARTS and CDDP were added to each cell line at final concentrations of 9.375 µM and 0.75 µg/ml or 46 µM and 1.0 µg/ml, respectively. The HNSCC cells (UM-SCC-23 and UM-SCC-81) were collected after 72 h and the levels of Rb and pRb were examined by western blotting. β-actin was used as a loading control. (B and C) Results are shown as relative values for the concentration of each band obtained by western blotting compared with (B) the untreated control and (C) pRb and Rb. Results are shown comparing the mean from three experiments. (D) Results are shown as relative values for the concentration of each band obtained by RT-qPCR compared with the untreated control. ARTS and CDDP were added to each cell line at final concentrations of 9.375 and 46 µM, respectively. Results show a comparison of data from three experiments. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. ARTS, artesunate; Rb, retinoblastoma protein; pRb, phosphorylated Rb; HNSCC, head and neck squamous cell carcinoma; CDDP, cisplatin; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: The drugs used were artemisinin (CAS RN ® : 63968-64-9; cat. no. A2118; Tokyo Chemical Industry Co., Ltd.), deoxyartemisinin (CAS RN ® : 72826-63-2; cat. no. D232150; Toronto Research Chemicals), dihydroartemisinin (CAS RN ® : 81496-82-4, cat. no. D3793; Tokyo Chemical Industry Co., Ltd.), artesunate (CAS RN ® : 88495-63-0, cat. no. A2191; Tokyo Chemical Industry Co., Ltd.), and cisplatin (Randa Injection; Nippon Kayaku Co., Ltd.).

Techniques: Western Blot, Control, Concentration Assay, Quantitative RT-PCR, Comparison, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

doi: 10.3892/or.2023.8591

Figure Lengend Snippet: Effects of ARTS on the expression of cell cycle-related molecules in the UM-SCC-23 cell line. (A) Western blotting was performed for CDK2, CDK4, CDK6, cyclin B1, cyclin D1 and cyclin E in UM-SCC-23 cells treated as in . (B) Results are shown as relative values for the concentration of each band obtained by western blotting compared with untreated controls. Results are shown comparing data from three experiments. *P<0.05. ARTS, artesunate; CDDP, cisplatin.

Article Snippet: The drugs used were artemisinin (CAS RN ® : 63968-64-9; cat. no. A2118; Tokyo Chemical Industry Co., Ltd.), deoxyartemisinin (CAS RN ® : 72826-63-2; cat. no. D232150; Toronto Research Chemicals), dihydroartemisinin (CAS RN ® : 81496-82-4, cat. no. D3793; Tokyo Chemical Industry Co., Ltd.), artesunate (CAS RN ® : 88495-63-0, cat. no. A2191; Tokyo Chemical Industry Co., Ltd.), and cisplatin (Randa Injection; Nippon Kayaku Co., Ltd.).

Techniques: Expressing, Western Blot, Concentration Assay

Journal: Oncology Reports

Article Title: Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

doi: 10.3892/or.2023.8591

Figure Lengend Snippet: Effect of ARTS on the expression of cell cycle-related molecules in the UM-SCC-81B cell line. (A) Western blotting was performed for CDK2, CDK4, CDK6, cyclin B1, cyclin D1 and cyclin E in UM-SCC-81B cells treated as in . (B) Results are shown as relative values for the concentration of each band obtained by western blotting compared with untreated controls. Results are shown comparing data from three experiments. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. ARTS, artesunate; CDDP, cisplatin.

Article Snippet: The drugs used were artemisinin (CAS RN ® : 63968-64-9; cat. no. A2118; Tokyo Chemical Industry Co., Ltd.), deoxyartemisinin (CAS RN ® : 72826-63-2; cat. no. D232150; Toronto Research Chemicals), dihydroartemisinin (CAS RN ® : 81496-82-4, cat. no. D3793; Tokyo Chemical Industry Co., Ltd.), artesunate (CAS RN ® : 88495-63-0, cat. no. A2191; Tokyo Chemical Industry Co., Ltd.), and cisplatin (Randa Injection; Nippon Kayaku Co., Ltd.).

Techniques: Expressing, Western Blot, Concentration Assay

Journal: Parasite

Article Title: Assessment of quantitative and semi-quantitative biological test methods of artesunate in vitro

doi: 10.1051/parasite/2022019

Figure Lengend Snippet: Parasite growth inhibition curve with artesunate concentration for the quantitative test. This curve resulted from the analysis of data sets from nine independent assays using a four-parameter nonlinear regression. Each concentration was tested in triplicate in each independent assay. The X -axis was transformed into logarithm to the base 10 to better visualize the linear portion of the curve. Data are presented as the mean ± SD ( n ≥ 9). FU = fluorescence unit; Log 10 = logarithm to the base 10; [AS] = artesunate concentration; nM = nanomolar; Red-dotted lines = indicate linearity interval.

Article Snippet: Artesunate (Chemical Abstract Service CAS number: 88495-63-0; IPCA #19003AS6RII [1 kg]) was purchased from IPCA Laboratories Limited (Mumbai, India) through Hepartex (Saint-Cloud, France).

Techniques: Inhibition, Concentration Assay, Transformation Assay, Fluorescence

Journal: Parasite

Article Title: Assessment of quantitative and semi-quantitative biological test methods of artesunate in vitro

doi: 10.1051/parasite/2022019

Figure Lengend Snippet: Validation data of the standard curve using controls samples of fixed artesunate concentrations.

Article Snippet: Artesunate (Chemical Abstract Service CAS number: 88495-63-0; IPCA #19003AS6RII [1 kg]) was purchased from IPCA Laboratories Limited (Mumbai, India) through Hepartex (Saint-Cloud, France).

Techniques:

Journal: Parasite

Article Title: Assessment of quantitative and semi-quantitative biological test methods of artesunate in vitro

doi: 10.1051/parasite/2022019

Figure Lengend Snippet: Standard curve of the quantitative test with the 95% confidence interval for artesunate controls samples . The equation of the curve (■) was y = −1831 x + 292150 ( r 2 = 0.9373). The antimalarial activities of the controls ( ), plotted against their respective expected concentrations (Q1 = 15 nM and Q2 = 30 nM), fell outside the 95% confidence interval of the curve. Data are presented as mean ± SD ( n ≥ 6). FU = fluorescence unit; AS = artesunate; nM = nanomolar.

Article Snippet: Artesunate (Chemical Abstract Service CAS number: 88495-63-0; IPCA #19003AS6RII [1 kg]) was purchased from IPCA Laboratories Limited (Mumbai, India) through Hepartex (Saint-Cloud, France).

Techniques: Fluorescence

Journal: Parasite

Article Title: Assessment of quantitative and semi-quantitative biological test methods of artesunate in vitro

doi: 10.1051/parasite/2022019

Figure Lengend Snippet: (a) Impact of KRB on Pf growth and (b) antimalarial activity of artesunate at 4 nM prepared in culture medium or 12.5% KRB in culture medium for the semi-quantitative test. KRB concentrations above 12.5% (v/v) significantly reduced parasites growth. All samples containing KRB were therefore diluted into culture medium before use to limit its impact. Each concentration was tested in triplicate in two independent assays. Data are presented as the mean ± SD ( n = 6). There was no statistical difference between the antimalarial activities of 4 nM artesunate solutions prepared in culture medium and 12.5% (v/v) KRB. Each sample was tested in triplicate in three independent assays. Data are presented as mean ± SD ( n = 9). KRB = Krebs ringer buffer; AS = artesunate; FU = fluorescence unit; ns: non-significant. Statistical analyses were performed with GraphPad Prism software, Mann–Whitney test, α = 5%, p -value (**) = 0.0022.

Article Snippet: Artesunate (Chemical Abstract Service CAS number: 88495-63-0; IPCA #19003AS6RII [1 kg]) was purchased from IPCA Laboratories Limited (Mumbai, India) through Hepartex (Saint-Cloud, France).

Techniques: Activity Assay, Concentration Assay, Fluorescence, Software, MANN-WHITNEY

Journal: Parasite

Article Title: Assessment of quantitative and semi-quantitative biological test methods of artesunate in vitro

doi: 10.1051/parasite/2022019

Figure Lengend Snippet: Antimalarial activity of the reference condition (AS 4 nM) and samples from (a) the permeation test using artesunate solution and (b) the permeation test using artesunate powder. Each sample was tested in triplicate. Data are presented as mean ± SD ( n = 3). AS = artesunate; FU = fluorescence unit; Green dotted-line = threshold value for maximum parasite growth (fluorescence of parasites incubated without artesunate and KRB during the semi-quantitative test); Red dotted-line = threshold value for minimum parasite growth (parasite fluorescence at the beginning of the test).

Article Snippet: Artesunate (Chemical Abstract Service CAS number: 88495-63-0; IPCA #19003AS6RII [1 kg]) was purchased from IPCA Laboratories Limited (Mumbai, India) through Hepartex (Saint-Cloud, France).

Techniques: Activity Assay, Fluorescence, Incubation