RNA-seq transcriptome analysis of breast cancer cell lines under shikonin treatment

Abstract

Shikonin is a naphthoquinone isolated from the dried root of Lithospermum erythrorhizon, an herb used in Chinese medicine. Although several studies have indicated that shikonin exhibits antitumor activity in breast cancer, the mechanism of action remains unclear. In the present study, we performed transcriptome analysis using RNA-seq and explored the mechanism of action of shikonin in regulating the growth of different types of breast cancer cells. The IC 50 of shikonin on MCF-7, SKBR-3 and MDA-MB-231 cells were 10.3 μΜ, 15.0 μΜ, 15.0 μΜ respectively. Our results also demonstrated that shikonin arrests the progression of cell cycle and induces apoptosis in MDA-MB-231 cells. Using RNAseq transcriptome analysis, we found 38 common genes that significantly express in different types of breast cancer cells under shikonin treatment. In particular, our results indicated that shikonin induces the expression of dual specificity phosphatase (DUSP)-1 and DUSP2 in both RNA and protein levels. In addition, shikonin also inhibits the phosphorylation of JNK and p38, the downstream signaling molecules of DUSP1 and DUSP2. Therefore, our results suggest that shikonin induces the expression of DUSP1 and DUSP2 which consequently switches off JNK and p38 MAPK pathways and causes cell cycle arrest and apoptosis in breast cancer cells.Breast cancer is one of the most common cancers and the second leading cause of cancer death among women in the United States 1 . One in eight women will be diagnosed with breast cancer in her lifetime. Approximately 70% of breast cancer patients are inoperable because of advanced tumor growth or bone metastasis 2 . Therefore, new strategies for the treatment of breast cancer are necessary. Many agents extracted from Traditional Chinese medicine (TCM) have been shown to possess anticancer activities and can be considered as alternative treatments for breast cancer 3 .Shikonin, a naphthoquinone isolated from the Chinese herbal plant Lithospermum erythrorhizon, has been used to treat a variety of inflammatory and infectious diseases 4 . Several biological and pharmacological actions of shikonin have been reported, including anti-inflammatory 5 , antibacterial 6 , antiviral 7 , and antioxidant 8 activities. In particular, shikonin has been shown to exert anticancer properties via different mechanisms on various

Shikonin and 4-hydroxytamoxifen synergistically inhibit the proliferation of breast cancer cells through activating apoptosis signaling pathway in vitro and in vivo

“…SK is a well-known active ingredient isolated from the root of a Chinese herbal medicine that has been used for thousands of years. Its extensive and remarkable antitumor activities, including its anti-breast [18], gastric [34], colon [35], glioma [36] effects, have attracted considerable attention. Breast cancer research findings show SK could inhibit estrogen-dependent tumor cell growth and promote the anti-estrogen effect of TAM, restoring its efficacy against cancer cells [19].…”

“…Shikonin (SK) is a natural naphthoquinone compound extracted from the roots of the Chinese herbal plant Lithospermum erythrorhizon, exhibiting powerful anticancer activities on various breast cancer cells [18]. This effect is accomplished through multiple pathways that disrupt ER recruitment [19], induce programmed cell death [20,21], and inhibit metalloprotease (MMP)-9 [22] and topoisomerase [23].…”

Shikonin inhibits migration and invasion of triple-negative breast cancer cells by suppressing epithelial-mesenchymal transition via miR-17-5p/PTEN/Akt pathway

“…Administration of shikonin and its derivatives at a high dose is verified to be safe and well-tolerated in animal models [27,28]. In breast cancer, there are several researches confirm that SHK could induce apoptosis and cell cycle arrest, inhibit drug resistances and enhance cytotoxicity of chemotherapeutic agents [29,30]. As shown in the present study, shikonin effectively inhibited the migration and invasion of TNBC cell line, indicating that the use of SHK might provide an effective way to reduce metastasis of TNBC.…”

Integration of RNA-seq and Ribosome Profiling to Characterize Transcriptional and Translational Estrogen Responses of Genes in Human Breast Cancer

“…High-throughput RNA sequencing (RNA-Seq) provides precise quanti cation of gene expression in living cells (Conesa et al, 2016) and yields accurate and comprehensive transcriptome pro les of tumors and normal cells, and helps to de ne and characterize gene-regulatory responses to treatments (Lin et al, 2018). Intriguingly, the correlation of mRNA and protein levels in mammalian cells is imperfect, indicating that the study of gene regulation solely from RNA-Seq data is inherently insu cient to understand the adaptive landscape of cells to stimuli (Haider & Pal, 2013).…”