2019AndrologiaOpen access

Effects of dihydroartemisinin on HSP70 expression in human prostate cancer PC‐3 cells

Jin Seon Kong, Shan‐Shan Li, Qi Ma, Lei Liu, Linjing Zheng

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Abstract

We aimed to evaluate the effects of dihydroartemisinin (DHA) on heat-shock protein 70 (HSP70) expression in human prostate cancer PC-3 cells and to examine the molecular mechanism. The viability of PC-3 cells following treatment with 25, 50, 100 and 200 μmol/L DHA for 48 hr was detected by flow cytometry and MTT assay. The expression of HSP70 mRNA was detected by RT-qPCR. The expression levels and locations of HSP70, caspase-3 and apoptosis-inducing factor (AIF) were detected with immunofluorescence assay. With 100 μmol/L HSP70 inhibitor quercetin as positive control and dimethyl sulphoxide (DMSO) as solvent control, the protein expressions of HSP70, apoptotic protease activating factor-1 (Apaf-1) and AIF were detected by Western blot. DHA promoted PC-3 cell apoptosis dose-dependently. With increasing DHA dose, the expression of HSP70 mRNA significantly decreased (p < 0.05). DHA did not change the location of HSP70 or AIF. Compared with control and DMSO groups, the expression of HSP70 protein significantly decreased, and those of Apaf-1, caspase-3 and AIF significantly increased following treatment with DHA and quercetin for 48 hr. In conclusion, DHA inhibits the expression of HSP70 and induces the apoptosis of PC-3 cells. The results provide valuable experimental evidence for prostate cancer therapies using DHA.

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What this paper is about

We aimed to evaluate the effects of dihydroartemisinin (DHA) on heat-shock protein 70 (HSP70) expression in human prostate cancer PC-3 cells and to examine the molecular mechanism. The viability of PC-3 cells following treatment with 25, 50, 100 and 200 μmol/L DHA for 48 hr was detected by flow cytometry and MTT assay. The expression of HSP70 mRNA was detected by RT-qPCR. The expression levels and locations of HSP70, caspase-3 and apoptosis-inducing factor (AIF) were detected with immunofluorescence assay. With 100 μmol/L HSP70 inhibitor quercetin as positive control and dimethyl sulphoxide (DMSO) as solvent control, the protein expressions of HSP70, apoptotic protease activating factor-1 (Apaf-1) and AIF were detected by Western blot. DHA promoted PC-3 cell apoptosis dose-dependently. With increasing DHA dose, the expression of HSP70 mRNA significantly decreased (p < 0.05). DHA did not change the location of HSP70 or AIF. Compared with control and DMSO groups, the expression of HSP70 protein significantly decreased, and those of Apaf-1, caspase-3 and AIF significantly increased following treatment with DHA and quercetin for 48 hr. In conclusion, DHA inhibits the expression of HSP70 and induces the apoptosis of PC-3 cells. The results provide valuable experimental evidence for prostate cancer therapies using DHA.

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Available abstract

We aimed to evaluate the effects of dihydroartemisinin (DHA) on heat-shock protein 70 (HSP70) expression in human prostate cancer PC-3 cells and to examine the molecular mechanism. The viability of PC-3 cells following treatment with 25, 50, 100 and 200 μmol/L DHA for 48 hr was detected by flow cytometry and MTT assay. The expression of HSP70 mRNA was detected by RT-qPCR. The expression levels and locations of HSP70, caspase-3 and apoptosis-inducing factor (AIF) were detected with immunofluorescence assay. With 100 μmol/L HSP70 inhibitor quercetin as positive control and dimethyl sulphoxide (DMSO) as solvent control, the protein expressions of HSP70, apoptotic protease activating factor-1 (Apaf-1) and AIF were detected by Western blot. DHA promoted PC-3 cell apoptosis dose-dependently. With increasing DHA dose, the expression of HSP70 mRNA significantly decreased (p < 0.05). DHA did not change the location of HSP70 or AIF. Compared with control and DMSO groups, the expression of HSP70 protein significantly decreased, and those of Apaf-1, caspase-3 and AIF significantly increased following treatment with DHA and quercetin for 48 hr. In conclusion, DHA inhibits the expression of HSP70 and induces the apoptosis of PC-3 cells. The results provide valuable experimental evidence for prostate cancer therapies using DHA.

Key concepts: Hsp70, Apoptosis, Flow cytometry, Dihydroartemisinin, Western blot, Molecular biology, Heat shock protein, MTT assay

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Effects of dihydroartemisinin on HSP70 expression in human prostate cancer PC‐3 cells — Research Paper | ScholarLens