**5.4. Skin cancer**

meat, obesity, smoking, lack of physical exercise, diabetes, inflammatory bowel disease, and some genetic and epigenetic alterations as: microsatellite instability, chromosomal instability, mutation of p53 gene is one of the familiar genetic changes in the development of colon cancer, and several others [58]. A very recent study published in 2017, used a mouse model, which treated them comparatively with azoxymethan (AOM)/dextran sodium sulfate (DSS) and anthocyanin-rich extract from bilberries for colon cancer development [59]. The anthocyanin extract administered to mice resulted in less inflammation of the colon and a reduced number of tumors than the control group. The formation and the growth of colorectal cancer in AOM/ DSS-treated Balb/c mice were prevented by anthocyanins. Another *in vivo* study investigated the chemopreventive activity of commercially available anthocyanin-rich extracts of bilberry, chokeberry, and grape prepared for the food industry [60] Colon cancer male rats treated with a colon carcinogen, azoxymethane, had multiple biomarkers investigated such as: the number and multiplicity of colonic aberrant crypt foci, colonic cell proliferation, urinary levels of oxidative DNA damage, and expression of cyclooxygenase (COX) genes. Compared to the control group, rats fed with different extracts showed several changes. In rats fed with bilberry, chokeberry, and grape extracts, the number of large aberrant crypt foci was reduced. The bilberry and chokeberry diet decreased the colonic cellular proliferation, and the grape and bilberry diets had lower COX-2 mRNA expression of gene. These results clearly support

Breast cancer is the second most common cause of cancer-associated mortalities in women. The American Cancer Society estimated that 60,290 new cases of breast carcinoma *in situ* were expected to be diagnosed among women in the United States during 2015 [61]. Understanding the biology of the human epidermal growth factor receptor 2 (HER2) helps with the classification, prognosis, and treatment of breast cancer because of the overexpression of HER2 identified in 15–20% cases. HER2 is involved in proliferation, angiogenesis, invasion, and metastasis [62]. A group of scientists have used injection of cyaniding-3-glucoside and peonidin-3-glucoside to evaluate the effect on the tumors of the rats used in the experiments [63]. Compared with the control group, the tumors treated with cyanidin-3-glucoside and peonidin-3-glucoside expressed lower levels of HER2 as well as Ki67, a proliferation marker, demonstrated with histopathological studies. Also, the treated tumors expressed higher levels of caspase 3, showing the apoptotic effect of the treatment. A recent published study [64] evaluated the cytotoxicity of an anthocyanin-rich extract from black rice (AEBR) on breast cancer cells *in vitro* and *in vivo*. This study demonstrated that black rice extract has promising roles against breast cancer. The oral administration of anthocyanin-rich extract from black rice (100 mg/kg/day) on nude mice bearing MDAMB-453 cell xenografts, significantly suppressed

Lung cancer emerged as the most common cancer worldwide, with 1.8 million new cases in 2012 [57]. The treatment and prevention for lung cancer remains scarce, comparing too

tumor growth and angiogenesis, as well as antagonized VEGF activity.

the chemopreventive activity of tested extracts.

80 Phytochemicals - Source of Antioxidants and Role in Disease Prevention

**5.2. Breast cancer**

**5.3. Lung cancer**

Malignant melanoma of skin accounted for 232,000 new cases, and the regions affected are largely those with white populations [57]. Melanoma skin cancer originates in melanocytes, specialized pigment-producing cells found in both the basal layer of the epidermis. Solar UVB radiation has been implicated as the main cause for skin cancer [67]. Early diagnosis is the key for curing this potentially deadly disease. Also prevention is playing a crucial role in spotting melanomas at earlier and more curable stages [68]. Biochemotherapy, the coadministration of traditional chemotherapeutic drugs and biological agents, show a higher response rate for patients than classical treatments that are based only on chemotherapy alone [69–72]. Most anticancer treatments are derived from natural resources such as marine, microbial, and botanical sources [72]. Natural supplements, a rich diet in antioxidants used as a complementary medication, become a common field of research in order to develop new products originating from natural sources with antioxidant and chemopreventive properties. The ability of anthocyanins to influence parameters of skin tumor development on mice was demonstrated in various studies. SKH-1 hairless mouse was used in order to investigate the photo-chemopreventive effect of delphinidin on UVBinduced biomarkers of skin cancer development [17]. After the treatment, the results suggest that delphinidin inhibited UVB-mediated oxidative stress and reduced DNA damage, thereby protecting the cells from UVB-induced apoptosis. The antitumor activity of the anthocyanins extract from *Fructus Sorbi aucupariae* on B-16 melanoma in C57BI/6 mice was also demonstrated [73]. The study revealed an increase in the counts of stromal progenitor cells in the tumor node and their accelerated maturation. The potentiation of the antimetastatic activity of the cytostatic was demonstrated as well. The inhibitory effects of mulberry anthocyanins on the metastasis of B16-F1 cells under noncytotoxic concentrations were investigated. The findings of the study have demonstrated that mulberry anthocyanins have strong anticancer effects by inhibiting the metastasis ability of B16-F1 cells. Further investigations revealed that the antimetastatic effect of these compounds was also evident in a C57BL/6 mice model.

#### **5.5. Prostate cancer**

Prostate cancer is the most common malignancy in men and affects most men over the age of 50 and also presents one of the main causes of mortality. For the *in vivo* study, athymic nude mice are used. To highlight the effects of anthocyanins on tumor growth *in vivo*, DH145 tumor xenograft have been established in these mice. The group of animals treated with anthocyanins received an oral dose of 8 mg/kg per day. The effects of treatment were analyzed every 4 weeks. In the first 4 weeks after incubation, the difference between the control and the treated group was insignificant. In the second set of analyses (8 weeks), the difference between the groups was very clear, the control group tumors being much bigger. These differences were observed until the end of the experiment, demonstrating the ability of anthocyanins to reduce tumor growth [74]. Another study has found that delphinidin is effective *in vitro* on PC3 cells and has determined whether these results are also visible in *in vivo* models. The delphinidin doses were not toxic to the animals because they did not lose weight and did not affect the amount of food they consumed. After measurements for 12 weeks, the differences between the tumors of the two groups (control and treated) each week were significant, suggesting an antiangiogenic effect on tumor cells. At the end of the experiment, tumors were extirpated and analyzed, where effects similar to *in vitro* studies were observed [75].

chemopreventive activity against colon cancer. The growth of colon-cancer-derived HT-29 and nontumorigenic colonic NCM460 cell lines exposed to semipurified anthocyanin-rich extracts (AREs) was monitored for up to 72 h. All extracts inhibited the growth of HT-29 cells, chokeberry extract being the most potent inhibitor. Most importantly, the growth of NCM460 cells was not inhibited at lower concentrations of all three extracts, illustrating better inhibition of colon cancer, as compared to nontumorigenic colon cells. Lately, another study [78] investigated and observed the effects of extracts from five cultivars of strawberries on the proliferation of colon cancer cells HT29 and breast cancer cells MCF-7. Using strawberry as a source of anthocyanins, they demonstrated that strawberry extracts decreased the prolifera-

Anthocyanins-Smart Molecules for Cancer Prevention http://dx.doi.org/10.5772/intechopen.79613 83

Human epidermal growth factor 2 (HER2) is a member of the epidermal growth factor receptor family which is overexpressed in breast cancer, and to study the *in vitro* effect of anthocyanins, the cell lines in breast cancer are usually HER2 positive; unfortunately, there are many other types of breast cancer that occur due to other causes. Regarding potential chemopreventive effects of anthocyanins, recently, it was demonstrated that black rice anthocyanins reduce the adhesion, migration, and invasion of HER2 MDA-MB-453 cells. The morphology of these cells was significantly altered, moving from a mesenchymal to an epithelial state. The western blot analysis shows an increase of the epithelial marker, E-cadherin, and decreased the expression of the mesenchymal markers, fibronectin and vimentin; this shows the effect that BRAC has on epithelial mesenchymal transition (EMT). EMT is a process by which epithelial cells lose their cell polarity and cell-cell adhesion, and gain migratory and invasive properties which occurs in the initiation of metastasis in cancer progression [4]. An important role in the metastasis of MDA-MB-453 cells is the focal adhesion kinase (FAK)-signaling pathway. FAK promotes the increased expression of transcription factors associated with EMT. The cells used in certain analysis were treated with Y15 (FAK inhibitor) that inhibits the autophosphorylation site of FAK. The study shows that BRAC has a similar effect to Y15, and also

Inhibitory effect of anthocyanins on the migration and invasion of lung cancer was also studied. A previous study reported that glycosylated cyanidins isolated from mulberry exerted a dose-dependent inhibitory effect on the migration and invasion of metastatic A549 human lung carcinoma cells. Their results showed that the applied treatments could decrease the expressions of matrix matalloprotinase-2 (MMP-2) and urokinase plasminogen activator (u-PA) in a dose-dependent manner and also enhance the expression of tissue inhibitor of matrix matalloprotinase-2 (TIMP-2) and plasminogen activator inhibitor (PAI). Moreover, Western blot analysis revealed that anthocyanins treatment to A549 cells inhibited the activation of c-Jun (p48) and NF-kB (p65). Further, another study using anthocyanins from fruits of *Vitis coignetiae Pulliat* (AIMs) reported their anticancer effects on lung cancer cells. AIMs inhibited the growth; migration and invasion of A549 cells; and also some proteins involved with cancer effects are inhibited. AIMs suppressed MMP-2 (gelatinase-A) and MMP-9 (gelatinase-B), both involved in the

BRAC decreases the activation and transduction of FAK signaling [79].

tion of two cell lines in a dose-dependent manner.

**6.2. Breast cancer**

**6.3. Lung cancer**

#### **5.6. Leukemia**

Acute myeloid leukemia is a hematological malignancy that has numerous causes such as chromosomal abnormalities and various gene mutations. Fifty years ago, this type of cancer was incurable, but now around 35–40% of the cases is treatable [85]. Mice Balc/c has been used to identify *in vivo* benefits of mulberry anthocyanins. Leukemia mice treated with anthocyanins had a higher survival rate than the untreated ones, this survival being correlated with the concentration of treatment. All leukemia-induced mice had the spleen and liver measured at autopsy, indicating splenomegaly and hepatomegaly. The size of these organs was significantly reduced for those treated compared to the control group. The organs were evaluated histopathologically as well and again the treated group had less infiltrated tissue with leukemic cells. Taken all this into consideration, we can say that mulberry anthocyanins can improve or eliminate the leukemic mice disorder [86].
