Group members Mrs. Prisna Pianjing Ms. Monta Thammasatta Ms. Kunnika Changwichan Mrs. Indra Gurung Ms. Davong Oumavong

is Hepatocellular carcinoma (HCC) Cancer arise from hepatocyte It is found at peak age years and commonly occur in men Major causes are Hepatitis viral infection and Aflatoxin B1 Its 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 hepatocarcinogenesis Chronic hepatitis and cirrhosis commonly occur in preneoplastic stage Hepatitis B core antigen (HBcAg),Hepatitis Bsurface antigen (HBsAg),HBx Hepatitis C virus DNA Adduct by AflatoxinB1 Combination

Diagnosis of HCC The 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

Surgical Resection,Transplantation -Non surgical Chemotherapy Prognosis of HCC Treatment of HCC Depends on the size of the tumor and the 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 detection Treatment is high cost and often recurrence Harmful infection and difficult to treat Exposure of risk factors are difficult to avoid That why we must focus on prevention

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

Prevention Therapy Chemo preventive agent - High Risk - Non-toxic - Altered Food - Molecular markers - Combination CPACTA -Cancer Patients -Toxicity acceptable - Molecular marker -Synthetic drugs -Combination with CPA CPA : Chemopreventive agents CTA : 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 B1 The enzyme conjugate Aflatoxin B1 for excreting out via urine is.. Glutathione S-transferase (GSTs) Aflatoxin-N-acetylcysteine

To enhance this reaction, GST must be activated Sulphoraphan and curcuminoid have high potency to induce phase II enzyme….…. We’ve known that these two substances can induce GST......How… Transcriptional control to phase II gene is achieve by the binding of Nrf2 (nuclear factor erythroid2-relate factor) to consensus activate antioxidant response element (ARE) Keap1 repress Nf2 but if it dissociate from Keap1 ARE will be activated and GST will be produced

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 compounds Isothiocyanate and Indoles

Phenethyl isothiocyanate (R = C6H5CH2CH2–) indoles isothionates Both groups have high affinity to induce Phase II enzyme and inhibit tumor proliferation.

Curcumin from Tumeric Curcumin is the main biological active compound of rhizome of tumeric It has unique ability to fit through aryl hydrocarbon receptor : estrogen receptor Curcumin is both strongly anti-inflammatory and strongly anti-oxidant. Reduce potency of chemotherapy drugs accumulate in liver if liberate at high dose Precaution:

Effect of chemopreventive on carcinogen-metabolizing enzyme compoundNAD(P)H Quinone reductase CD(μM) QR induction IC 50 (μM) Cyp1A Inhibition IC 50 (μM) Inhibition of ODC induction IC 50 (μM) Curcumin PEITC0.93.2>59.5 Vitamin C>50 >5>10 Source: Gerhauser, C and et.al. 2002

Mixing of curcumin with glucosinolate compound There 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 result Of induce apoptosis and inhibit tumor signaling We expect that using these two compound in one dose Will give the same result in HCC

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

Testing method for this assumption… Gene-reporter method as high throughput technique By using Luciferase assay to quantify efficiency In vitro….. 1.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 concentrationzinc and oltipraz 5. Lyses cell and measure luciferase activities

Plasmid construction

Premalignant cell prevention Enhance apoptosis&Inhibit of tumor promoters cascade Focus on cell survival signaling pathway ERK, MAPK,STAT and NfKB are in our focus

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

Expectation…… Getting 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 broccoli might be the the better alternative way to prevention Than using single compound.

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