Abstract:
Background: Breast cancer is the most common type of invasive cancer in women in their forties and fifties. Recent evidence suggests that JAK2/STAT3 signaling is constitutively active in breast cancer. Previous studies suggest that plant extracts, including Salvia Officinalis, have strong cytotoxic effects on breast cancer cells. The differential expression of miRNAs is also strongly linked to cancer initiation and progression.
Aim: In the current study, we hypothesize that S. Officinalis extract suppresses JAK2 expression and has strong anticancer potential in MCF7 breast cancer cells in vitro.
Methods: GC-MS analysis showed the presence of flavonoids in Salvia officinalis Extract. The cytotoxicity of S. Officinalis was compared to Cisplatin on human breast (MCF-7) cells. Bioinformatic analysis was performed to detect the link between JAK2 and different microRNAs. qPCR assessment of microRNAs and JAK2 , BAX, Bcl-xL and BIRC5 mRNAs was performed. miR-216a-5p was overexpressed in MCF7 cells to test its anticancer potential.
Results: GC-MS analysis showed the presence of anticancer and antioxidant compounds (Linolein and Apigenin). S. Officinalis extract reduced cell proliferation of MCF-7 cells with an IC50 range from 5.123 to 6.345 mg/mL (p<0.0001) compared to cisplatin (IC50 =20 ug/ul). S. Officinalis was also safe for the human skin fibroblast, suggesting that S. Officinalis has anticancer activity and is less harmful to normal cells (p<0.0001). Morphological assessment of MCF-7 cells showed that untreated cells maintained their epithelial morphological shape, while those treated with S. Officinalis displayed morphological changes consistent with apoptosis. Bioinformatic analysis revealed that JAK2 contains two theoretical binding sites of miR-101, miR-216, and miR-204 in its 3` UTR. qPCR revealed that three miRs (miR-101, miR-216a-5p & miR-204-5p) were low expressed in breast cancer cell lines than in normal cell lines (P=0.0022). S. Officinalis and cisplatin reduced the expression of miR-101-5p (p<0.0001). While S. Officinalis reduced the expression of miR-216 and miR-204, Cisplatin, on the other hand, increased their expression (p<0.005). Also, qPCR showed that S. Officinalis and miR-216a-5p mimics significantly reduced JAK2 mRNA expression (p<0.0001). Both S. Officinalis and miR-216a-5p increased the expression of BAX and reduced the expression of Bcl-xL and BIRC5.
Conclusions:
S. Officinalis has significant anticancer potential mediated through the increased expression of BAX and reduced expression of JAK2, Bcl-xL, and BIRC5 mRNAs. As well as the reduced expression of miR-101, miR-216, and miR-204.
Keywords: Salvia Officinalis, miR-101-5p, miR-216a-5p & miR-204-5p, JAK2, BAX, Bcl-xL, BIRC5
Audience Take Away Notes:
- To explain the importance of genetics in understanding the development of breast cancer
- Using different types of treatments to see to what extent miRNA and plant extract can be a potential therapeutic role for breast cancer
- Illustrate the different laboratory methods that were used such as transfection
