Tumor regression after systemic administration of tocotrienol entrapped in tumor-targeted vesicles

Fu JY, Blatchford DR, Tetley L, Dufès C.

J Control Release. 2009 Dec 3;140(2):95-9. Epub 2009 Aug 23.

The therapeutic potential of tocotrienol, an extract of vitamin E with anti-cancer properties, is hampered by its failure to specifically reach tumors after intravenous administration, without secondary effects on normal tissues. We hypothesize that the encapsulation of tocotrienol-rich fraction (TRF) within vesicles bearing transferrin, whose receptors are overexpressed on many cancer cells, could result in a selective delivery to tumors after intravenous administration. The objectives of this study are therefore to prepare and characterize transferrin-targeted vesicles encapsulating TRF, and to evaluate their therapeutic efficacy in vitro and in vivo. The entrapment of TRF in transferrin-bearing vesicles led to a 3-fold higher TRF uptake and more than 100-fold improved cytotoxicity in A431 (epidermoid carcinoma), T98G (glioblastoma) and A2780 (ovarian carcinoma) cell lines compared to TRF solution. The intravenous administration of TRF encapsulated in transferrin-bearing vesicles led to tumor regression and improvement of animal survival in a murine xenograft model, contrary to that observed with controls. The treatment was well tolerated by the animals. This work corresponds to the first preparation of a tumor-targeted delivery system able to encapsulate tocotrienol. Our findings show that TRF encapsulated in transferrin-bearing vesicles is a highly promising therapeutic system, leading to tumor regression after intravenous administration without visible toxicity.

BACKGROUND: Previous studies have shown that nicotine enhances oxidative DNA damage and leads to increased lipid peroxidation, which affects embryo development. The present study investigated the effect of daily supplementation of gamma-tocotrienol on oocytes of nicotine-treated mice.

MATERIAL/METHODS: Immature female mice (18-25 g) were divided into three groups. For 30 days, group A (control group) received saline (0.2 ml/day s.c.), group B nicotine (5 mg/kg/day s.c. in saline), and group C nicotine with gamma-tocotrienol (60 mg/kg/day p.o.). The animals were superovulated following these schedules.

RESULTS: Scanning electron microscopy (SEM) showed that the nicotine-treated oocytes appeared nonspherical with rough surface and the zona pellucida (zp) was torn and became irregular. Supplementation with gamma-tocotrienol in the nicotine-treated mice retained the spherical shape of the oocytes with intact zp; however, the surfaces of the oocytes remained irregular and rough. Transmission electron microscopy (TEM) following chronic nicotine treatment showed loosening of the boundary and tearing of the zp. The perivitelline space was also widened. The cytoplasm of the oocytes retained abundant rough endoplasmic reticulum (rER) with numerous vesicles. Mitochondria were highly dense, with no cristae. The administration of gamma-tocotrienol partially reduced the detrimental effects of nicotine by retaining the smooth boundary of the zp with the tight perivitelline space. There was less rER with no visible vesicle and a lower amount of dense mitochondrial matrix.

CONCLUSIONS: This study documented that chronic nicotine treatment adversely affects the ultrastructure of oocytes, while gamma-tocotrienol treatment at least minimizes the nicotine-induced damage to oocytes.

Modulation of melanin synthesis and its gene expression in skin melanocytes by palm tocotrienol rich fraction

Suzana Makpol, Nur Nadia Mohd Arifin, Zahariah Ismail, Chua Kien Hui, Yasmin Anum Mohd Yusof and Wan Zurinah Wan Ngah

African Journal of Biochemistry Research Vol.3 (12), pp. 385-392, December, 2009

Melanin is the pigment that determines skin color. Melanin synthesis is catalysed by the enzyme tyrosinase and is controlled by TYR, TYRP1 and TYRP2 genes. The objective of this study was to evaluate the anti pigmentation property of palm tocotrienol rich fraction by determining melanin synthesis and expression of genes involved in its regulation in skin melanocytes. Palm tocotrienol rich fraction (TRF) which contains 75% a-tocotrienol and 25% tocopherol was used to inhibit melanin synthesis which was determined by determining melanin level and tyrosinase enzyme activity. Expression of TYR, TYRP1 and TYRP2 genes was determined by quantitative real time reverse transcriptase polymerase chain reaction (real time RT-PCR). Primary culture of skin melanocytes was divided into two groups; untreated control and cells that were treated with 500 μg/ml tocotrienol rich fraction for 24 h. Our results showed that there was a reduction in tyrosinase activity and melanin content in melanocytes treated with tocotrienol rich fraction compared to control (p < 0.05). Expression of TYRP2 gene in melanocytes treated with tocotrienol rich fraction was also decreased (p < 0.05) compared to control. In conclusion, palm tocotrienol rich fraction has an anti pigmentation property that inhibit melanin synthesis by inhibiting tyrosinase activity and down regulating TYRP2 gene expression.