Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology,...

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© Kimmo Rumpunen SLU Balsgård Anticancer properties of sea buckthorn extracts and variation in content of ursolic acid among sea buckthorn cultivars grown in Sweden Kimmo Rumpunen Department of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU)

Transcript of Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology,...

Page 1: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLU Balsgård

Anticancer properties of sea buckthornextracts

and variation in content of ursolic acid among sea buckthorncultivars grown in Sweden

Kimmo RumpunenDepartment of Plant Breeding and Biotechnology, Balsgård,

Swedish University of Agricultural Sciences (SLU)

Page 2: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

Balsgård – division ofhorticultural plantbreeding

• Research on genetics and breeding offruits and berries

• Plant breeding of apple (since 1945),black currants (since 1950) and seabuckthorn (since 1985)

• Vast collections of apple, pear, plum,cherries, chaenomeles, currants, seabuckthorn, rose hips, cowberries

• Centre of innovative beverages(2010)

© Kimmo Rumpunen SLU Balsgård

Page 3: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLUBalsgård

• Adaptation to local climate (hardiness, early ripening)• Sweet taste (high sugar, low acidity)• Absence of rancidity at full (over) maturity• Consistent and high yield• Suitability for cut and freeze harvesting

Breeding andselection goals forsea buckthorn inSweden

Page 4: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

New selections

© Kimmo Rumpunen SLU Balsgård

BHi 10726

Strong growth, green foliageFew medium thornsCircular to oblong fruits of mediumsizeDense fruit clustersDistinctive aroma and medium acidityEarly to medium ripeningVery rich yield

BHi 32415

Weak growth, gray foliageFew long sharp thornsOblong to elliptic fruits of medium sizeSemi sweet fruits at full ripenessNo rancidity, dense fruit clustersEarly to medium ripeningRich yield

Page 5: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLU Balsgård

BHi 727115

Medium growth, green-gray foliage, fewthorns of medium sizeOvate to oblong fruits of medium sizeEarly ripeningSemi sweet at full maturity, no ranciditySemi dense fruit clustersRich yield

BHi 72587

Weak to medium growth, green-grayfoliage, few thorns of medium sizeVery large ovate to oblong fruitsVery early ripeningLong fruit stalksEasy to pick by handMedium yield

Page 6: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLU Balsgård

Antiproliferative effects of seabuckthorn (Hippophae rhamnoides L.)

extracts on human colon and livercancer cell lines

Carl Grey a,*, Cecilia Widen b, Patrick Adlercreutz a, Kimmo Rumpunen b,Rui-Dong Duan ca Department of Biotechnology, Lund University, Swedenb Department of Plant Breeding and Biotechnology Balsga°rd, SwedishUniversity of Agricultural Sciences, Swedenc Gastroenterology and Nutrition Laboratory, Biomedical Center, LundUniversity, Sweden

Participants

Publication

Food Chemistry 120 (2010) 1004–1010

Page 7: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLU Balsgård

• (1) to compare the antiproliferative and apoptotic effects ofdifferent sea buckthorn extracts on human colon cancercells (the Caco-2 cell line) and human liver cancer cells (theHep G2 cell line).

• (2) to determine whether the antiproliferative activities ofsea buckthorn extracts are associated with the content ofdifferent bioactive compounds.

Objectives

Page 8: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLU Balsgård

• Decreased proliferation of colon cancer and breast cancer cells.Correlation with vitamin C and carotenoids. (Olsson et al. J AgrFood Chem 2004)

• Quercetin, kaempherol and myricetin induced apoptosis ofleukemia cells as well as decreased cell proliferation. (Hibasami etal. Int J Mol Med 2005)

• Antiproliferative and apoptotic effects on forestomach and skincancer. (Padmavathi et al. Nutr Cancer 2005)

• Isorhamnetin can cause apoptosis in liver cancer cells. (Teng et al.Pharmacol Res 2006)

• Inhibition of stomach, prostate, intestine and breast cancer celllines by sea buckthorn juice. (Boivin et al. Anticancer Res 2007)

Background: Some previous anti-cancer studies on sea buckthorn

Page 9: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLU Balsgård

Plant material:• BHi 10726 (for sequential and solvent mixture extraction)• 24 cultivars and advanced selections (for screening of ursolic acid)

Cells:• Colon cancer cells (Caco-2)• Liver cancer cells (Hep G2)

Extraction procedures:• Sequential extraktion successively using n-heptane, ethyl acetate,

96% ethanol, and water.• Extraction by a solvent mixture of ethanol and acidified water (1:1).

Materials and methods

Page 10: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLU Balsgård

Cell analyses:• Cell viability• Cell proliferation• Cell apoptosis

Chemical analyses:• Total phenols• Phenolic compounds (rutin, isorhamnetin-3-O-rutinoside,

kaempferol-3-glucoside, isorhamnetin-3-O-rutinoside andproanthocyanidins)

• Ursolic acid

Materials and methods

Page 11: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

Results: Effects of different extractsof sea buckthorn on colon cancer cells

© Kimmo Rumpunen SLU Balsgård

Page 12: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

Results: Effects of different extractsof sea buckthorn on liver cancer cells

© Kimmo Rumpunen SLU Balsgård

Page 13: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

Results: Contents of differentsequential extracts and ethanol:water

extracts of sea buckthorn

© Kimmo Rumpunen SLU Balsgård

Page 14: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

Results: Ursolic acidin different sea

buckthorn cultivarsand selections

© Kimmo Rumpunen SLU Balsgård

Average: 2400 µg/g DW

Page 15: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

Results: Inhibition of cancer cellproliferation by ethyl acetate extracts of

different sea buckthorn cultivars

© Kimmo Rumpunen SLU Balsgård

Colon cancer cells Liver cancer cells

Page 16: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

Results: Composition of ethyl acetateextracts of different sea buckthorn

cultivars

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Page 17: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

Conclusions

• The strongest inhibitory effect was found in ethyl acetate extractfor colon cancer cells and in the ethanol:water extract for livercancer cells.

• The antiproliferative effects were in both cases dose-dependentand were in the case of the ethyl acetate extract associated withan increase in apoptosis.

• The ethyl acetate fraction contained high levels of ursolic acid andrather low amounts of phenols.

• The ethanol:water extracts contained high levels of phenols, butlittle ursolic acid.

• The dose dependent effects observed were not solely caused by asingle component and different extracts were most efficient fordifferent cells. This points out the advantage of using wholeextracts rather than isolated compounds when evaluating thephysiological relevance of fruits and berries.

© Kimmo Rumpunen SLU Balsgård

Page 18: Anticancer properties of sea buckthorn extractsDepartment of Plant Breeding and Biotechnology, Balsgård, Swedish University of Agricultural Sciences (SLU) Balsgård – division of

© Kimmo Rumpunen SLU Balsgård