Group membersMrs. Prisna PianjingMs. Monta ThammasattaMs. Kunnika ChangwichanMrs. Indra GurungMs. Davong Oumavong

http://www.healingdaily.com/conditions/cirrhosis

Hepatocellular carcinoma (HCC)Cancer arise from hepatocyteIt is found at peak age 50-70 years and commonly occur in menMajor causes are Hepatitis viral infection and Aflatoxin B1Its incidence is highest in sub-Saharan Africa, China, southern Asia, and Japan. HCC is the fifth leading cause of cancer mortality in the world

Sign and symptoms- Loss of appetite and weight

Abdominal pain, especially in the upper right part of the abdomen, that may extend into back and shoulder

- Nausea and vomiting

- General weakness and fatigue

- An enlarged liver

- Abdominal swelling (ascites)

A yellow discoloration of skin and the whites of eyes (jaundice)

Mechanism of hepatocarcinogenesisChronic hepatitis and cirrhosis commonly occur in preneoplastic stageHepatitis B core antigen (HBcAg),Hepatitis Bsurface antigen (HBsAg),HBxHepatitis C virusDNA Adduct by AflatoxinB1Combination

Diagnosis of HCCThe prognosis for patients with HCC after diagnosis is poor.The early detection of HCC might therefore be a mean to improve survival prospect.Done by basic laboratory evaluation and conducts every 6-12 months in patients at risk. Ultrasonography, MRI and CT scan can be used as detection for HCC associate with blood evaluation. How to detect HCC earlier.

Development of Hepatocellular carcinoma

SurgicalResection,TransplantationNon surgicalChemotherapy Prognosis of HCCTreatment of HCCDepends on the size of the tumor andthe extent of liver function impairment The life expectancy of HCC patients is hard to predict because of the high possibility of postoperative recurrence.

Poor prognosis is result from many factors:Poor early detectionTreatment is high cost and often recurrenceHarmful infection and difficult to treatExposure of risk factors are difficult to avoidThat why we must focus on prevention

Chemoprevention & Chemo interventionThe most important public health and medical practice is prevention of cancerChemopreventive agents are any and all natural or synthetic compound that can Inhibit, suppress, or reverse the development and progression of cancerChemo preventive agents can be administered as dietarycomponent in food dietary supplements or pharmaceuticals preparations. Prevent is better than cure

PreventionTherapyChemo preventiveagent High Risk Non-toxic Altered Food Molecular markers CombinationCPACTACancer PatientsToxicity acceptable Molecular markerSynthetic drugsCombination with CPACPA : Chemopreventive agentsCTA : Chemotherapeutic agents

What are we going to do.Stimulate detoxificatin aflatoxinB1 removed

Enhance production of Phase II enzyme Which way we can do in case of HCC.- Enhance detoxification- Prevent premalignant cell- Early detection

Phase II enzyme for removal Aflatoxin B1The enzyme conjugate Aflatoxin B1 for excretingout via urine is.. Glutathione S-transferase (GSTs)

Aflatoxin-N-acetylcysteine

To enhance this reaction, GST must be activatedSulphoraphan and curcuminoid have high potencyto induce phase II enzyme.Weve known that these two substances caninduce GST......HowTranscriptional control to phase II gene is achieve bythe binding of Nrf2 (nuclear factor erythroid2-relate factor)to consensus activate antioxidant response element (ARE)Keap1 repress Nf2 but if it dissociate from Keap1ARE will be activated and GST will be produced

www.ptinr.com/data/articlefiles/39-sprout.jpg www.communitygarden.org.au/.../root/tumeric.jpg Broccoli : A cruciferous vegetable that has high potency in prevention of cancer.How!!It contains a large numbers of phytochemical:The most significant is Glucosinolate It is a precursor of 2 groups of active compoundsIsothiocyanate and Indoles

www.rsc.org/images/soundbite3-400_tcm18-50620.jpg

Phenethyl isothiocyanate (R = C6H5CH2CH2) indoles isothionates Both groups have high affinity toinduce Phase II enzyme andinhibit tumor proliferation.

www.samispices.com/images/turmericimg.gif Curcumin from TumericCurcumin is the main biological active compound ofrhizome of tumericIt has unique ability to fit through aryl hydrocarbonreceptor : estrogen receptorCurcumin is both strongly anti-inflammatory andstrongly anti-oxidant. Reduce potency of chemotherapy drugsaccumulate in liver if liberate at high dosePrecaution:

Effect of chemopreventive on carcinogen-metabolizing enzymeSource: Gerhauser, C and et.al. 2002

compoundNAD(P)HQuinone reductaseCD(M)QR inductionIC50 (M)Cyp1AInhibitionIC50 (M)Inhibition of ODC inductionIC50 (M)Curcumin2.79.70.743.7PEITC0.93.2>59.5Vitamin C>50>50>5>10

Mixing of curcumin with glucosinolate compoundThere are many researches indicate that using the combination of curcumain and glucosinolate give higher advantage to treat prostate cancer

Using both substances at low dose give good resultOf induce apoptosis and inhibit tumor signalingWe expect that using these two compound in one doseWill give the same result in HCC

2 mechanismsPhosphorelate Nrf2 by MAPK Direct interaction of electrophilic agents with reactive thiol residues .

Testing method for this assumptionGene-reporter method as high throughput techniqueBy using Luciferase assay to quantify efficiencyIn vitro..Plasmid construct by using pGL-3 promoter vector consists of firefly luciferase (luc) gene driven by simian virus 40 (SV40) 2. This vector was insert with ARE transcriptional enhancer sequence 3. Transfect the vector into cell (primary liver cell & HepG2)4. Incubate cells with broccoli and curcumin extract and control substances (zinc and oltipraz), varies time and concentration5. Lyses cell and measure luciferase activities

Plasmid construction

Premalignant cell preventionEnhance apoptosis&Inhibit of tumor promoters cascadeFocus on cell survival signaling pathwayERK, MAPK ,STAT and NfKB are in our focus

Cell line ; same as gene reporter methodWestern blottingTesting substances- mixing of broccoli extract and curcumin- Test single compound- Positive control are piroxicam Material and method

ExpectationGetting the effective concentration from in vitro experiment that can be used further in vivo, clinical trial and products The using of combination of curcumin and broccolimight be the the better alternative way to prevention Than using single compound.

Zinc was examined in cultured retinal pigment epithelialcells that it can induce GST enzyme by using Luciferase method.(Ha, Khoi-Nguyen., et. al. 2006) Oltipraz was evaluated as chemopreventive agentin rat and can inhibit preneoplastic cell in liverafter expose to AFB1.(Roebuck,B D., et al. 2003)