HPLC phenolic profile and induction of apoptosis by Linum usitatissimum extract in LNCaP cells by caspase3 and Bax pathways

Abstract

Abstract Linum usitatissimum is a candidate as a remedy to treat prostate problems in some folklore medicines. In this study, we have reported the phenolic and flavonoid constituents, antioxidant activity, and potential of the plant extract against prostate cancer cells. The phenolic and flavonoid compound profile of the extract were established using HPLC analysis. While the total phenolic and flavonoid content (TPC and TFC) were analyzed using classic methods. The antioxidant activity of the extract was also evaluated. MTT assay and flow cytometry technique was used to evaluate antiproliferation activity and induction apoptosis of the plant extract on prostate cancer cells of LNCaP. We also evaluated the gene expression of Bax and caspase-3 using the real-time qPCR assay. HPLC result revealed that L. usitatissimum extract (LUE) was rich in phenolic acids such as gallic, ferulic, and vanillic acid with the amount of 3.56, 2.12, 1.24 μg/g extract respectively. 383.4 mg GAE/g and 47.1 mgRuE/g were calculated for total phenolic and flavonoid content. LUE exhibited radical scavenging activity with IC50 = 19.3 ± 1.1 µg/mL. LUE chelated ferrous ions with IC50 = 121.1 ± 1.3 µg/mL. LUE showed anti-proliferative activity on LNCaP cells with the IC50 values of 8.3, 6.3, and 5.4 μg/mL after 24, 48, and 72 h treatment. LUE also increased cell mortality by inducing apoptosis (15.3–29.8%). The real-time qPCR results exhibited an increase in gene expression of Bax and caspase-3. Our in vitro study demonstrates that L. usitatissimum can be considered as an effective agent to inhibit the growth and invasion the human prostate cancer cells.

Keywords

• Linum usitatissimum
• Phenolic profile
• Prostate cancer
• Induction apoptosis
• Bax
• Caspase-3