Cytotoxicity and Apoptotic Inducibility of Vitex agnus-castus Fruit Extract in Cultured Human Normal and Cancer Cells and Effect on Growth.

Abstract

Vitex agnus-castus is a shrub belonging to the Verbenaceae family, which grows naturally and is common in the Middle East and southern Europe. Its ripened fruits have been used as a folk medicine for a long time for alleviation and/or improvement of symptoms from obstetric and gynecological diseases. Hobbs, in "Vitex: The Women's Herb", described Vitex as an effective remedy for emmeniopathy, including amenorrhea, oligomenorrhea, and menorrhagia, and that due to estrogen hypersecretion before menstruation in premenstrual tension, long-term use of V. agnus-castus reduced those symptoms and prevented relapse of the disease. 1)It also appears that the function of the corpus luteum in corpus luteum deficiency syndrome improved with V. agnus-castus due to the crinogenic effect of luteinizing hormones.Hirobe et al. reported that an antitumor effect results from the extract of ripened Israeli-grown V. agnus-castus fruits (Vitex extract) on the Chinese hamster lung carcinoma cells line V-79, and that Vitex extract has high antitumor activity. 2)Furthermore, analysis of the components of Vitex extract showed four types of new flavonoid compound and four known flavonoids.3) We investigated the cytotoxicity of Vitex extract against a normal human cell line, human uterine cervical fibroblasts (HCF), and the cancer cell lines breast carcinoma (MCF-7) and ovarian cancer (SKOV 3). Using these three cell lines, we found that the cytotoxic activity of Vitex extract may be attributed to the growth activity of the respective cell, and demonstrated the possibility that the cytotoxicity is related to the cell cycle stage. 4)To investigate the antitumor effects of Vitex extract in more detail, we examined the cytotoxicity of Vitex extract against two noncancerous and six cancerous human cell lines. The cytotoxic activity of Vitex extract was confirmed to depend on the growth rate of the cells, the cytotoxic cell death of cells with relatively low growth activity occurs through apoptosis, and apoptotic cell death results from an increase in intracellular oxidation. MATERIALS AND METHODS Cells and Media The breast carcinoma (MCF-7)5) and gastric signet ring carcinoma (KATO III) 6) cell lines were described previously. Cervical carcinoma (SKG-3a), colon carcinoma (COLO 201), and small cell lung carcinoma (Lu-134-A-H) cell lines were provided by the JCRB Cell Bank (Tokyo, Japan). The ovarian cancer (SK-OV-3) cell line was provided by the American Type Culture Collection (Rockville, MD, U.S.A.). Fetal fibroblasts (HE-21) were obtained from the Research Institute for Functional Peptides (Yamagata, Japan). HCF was prepared as previously described.7) Pyrrolidine dithiocarbamate (PDTC) and N-acetyl-L-cysteine (NAC) were purchased from Dojindo Laboratories and Wako Pure Chemical Industries, respectively. Preparing Ethanol Extract from Dried Ripened V. agnus-castus Fruits Dried, ripened, Israeli-grown V. agnuscastus fruits were triturated, and a crude extract was prepared from the triturate (1 g) with ethanol (10 ml) under reflux for 2 h. After...

Cytocidal Effects of Polyphenolic Compounds, Alone or in Combination with, Anticancer Drugs Against Cancer Cells: Potential Future Application of the Combinatory Therapy

“…Of those, we have demonstrated that Vitex exhibits cytotoxic activities against various types of solid tumor cells, such as KATO-III (a human gastric signet ring carcinoma cell line), COLO 201 (a human colon adenocarcinoma cell line), MCF-7 (a human breast carcinoma cell line) [52]. More interestingly, no apparent cytotoxicity was observed in non-tumor cells, such as human uterine cervical canal fibroblast (HCF) and human embryo fibroblast (HE-21) when treated with concentrations showing significant cytotoxicity in tumor cells, suggesting a selective cytotoxic activity against tumor cells [52]. We further demonstrated that Vitex induced apoptosis accompanied by an accumulation of intracellular ROS along with the decrease in the levels of intracellular GSH in KATO-III cells [22].…”

Recommendations on Herbs and Herbal Formula in Cancer Prevention

“…It is widely known to cause homeopathic regulation of female endocrine system to ameliorate symptoms of PMS, amenorrhea, infertility, menopause due to estrogenic effects of phytoesterogens present in vitex [26], blocking production of prolactin from pituitary, shortening luteal phase and antagonizing hormonal imbalance(low progesterone synthesis in luteal phase) [27]. Its inducing cell death (tumoricidal) action is established in ovarian, cervical, breast, gastric, colon, lung cancer cells via oxidative stress related induction of pro-apoptotic caspase 3,8,9 hydroxy oxidase (a reduction in BCL-2, BCL-XL and Bid protein, increased Bad gene expression and induced DNA fragmentation [28,29]. However, it showed side effects of nausea, headache, rashes.…”

Effect of Vitex agnus Extract on MNU Induced Mammary Tumor of Sprague Dawley Rats

“…7 Further studies on it showed that ethanolic extract from dried ripe VAC fruits displays cytotoxic activity against six human cell lines from different cancer tissue origins. 8 Studies for antioxidant activity in the past have shown that decreased GSH levels are indicator of cell viability along-with increased MDA levels showing marked cell proliferation in tumor tissues whereas decreased levels of the same are indicator improved oxidative status of the tissue. From the recent literature, it is evident that ethanolic fruit extract of the VAC has shown some anticancer effects in, in-vitro studies and there is no recent data suggesting the anticancer property in vivo.…”