一下 ROBERT 論文討論會台大生科 (secondary paper discussion in NTULS )

8/8/2019 ROBERT (secondary paper discussion in NTULS )

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(Abstract)


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(Abstract-1)


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(Abstract-2)

!!!


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HSC-3OEC-M1OC3

1.

2.

3.

4.G1G2/M

5.Caspasebcl-2JNK

(Abstract-3)


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(Abstract-4)

Z-VAD(Caspase)

:

caspase

bcl-2JNK(MAPK)


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;caspasebcl-2(mitogen-activatedprotein kinase)

(Abstract-5)


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INTRODUCTION


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INTRODUCTION-1

Therearehigh incidences of humanheadand

neck squamous cellcarcinoma (HNSCC) insouth

Asia ,which include taiwan

Therisk of oralcavity cancer (one kind of the

HNSCC) is increasedby smokingoverdrinking ofalcohol and,especially in taiwan,arecachewing!

To dealwith it ,doctorusesurgeryradiotherapy

andchemotherapy,and they arenot satisfactory

due to many sideeffects


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INTRODUCTION-2

AFTER ALL,

THEY ARE ALL

TOXIC !!!!


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INTRODUCTION-3

Incontrast,paclitaxel isa naturalproduct

extracted from thebark ofpacific yew

(Taxus breviforlia)

It disrupt thedynamic of microtubules,which

inducesapoptosis (apts) andarresting cellat

G2/M phase

But the mechanism iselusive !!!


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INTRODUCTION-4

Youshouldnt expect

me to explainapts.

Again


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INTRODUCTION-5

Letsreview:

What are thecharacteristic ofapts??

How many right answers do you get ???Then,we are about to tell more in molecular

view !!

1. Cellarroundup (cytoplasmic contraction)

2. PM blebbing

3. DNA fragmetation

4. Chromatincondension


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INTRODUCTION-6

Thereare three majorpathway forapts

Caspase pathwaybcl-2 pathwayMAPKpathway.

They got signal from receptor on PM (mostly),triggering theapts by activating protease, Dnase,

inhibiting celldivisionand more.


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Caspasepathway

Signal from receptor on

PM or mt.

Caspase activation

DnaseDNA repair

enzymedegradation

(PARP cleavage)

related to otherpathway

INTRODUCTION-7


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There are two main pathwaysin Caspase-induced-apoptosis---death recptor(asshownbefore)and mitochondrial pathways.

Theyare activatedbycaspase -8,9respectively.

Andcaspase-3,6,7are downstream effectorinthe pathway

INTRODUCTION-7


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INTRODUCTION-8

Bcl-2 pathway


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INTRODUCTION-9

MAPK pathway(NO PROPER FIGURE)

GPCR on PM

Stimulate MAPKS,consist ofthreesub family: ERK,JNK,P-

38

Promoteapoptosis

In many mannerslikecellcycle

control,cellproliferation

Slowdown.


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In sum !!!


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Methodand Result


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Method-Cellculture

Skip !!!

Do it once inrealand youwill know much more than I

canexplain xdd


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Method-Morphology study

each 96-wellplatecontain 1*104 celland 100 ul of

serum

Each threecelllines 0(AS??),50,500nM paclitaxel

And incubate them for 24 and 48 hr

Observe them under LM andphotograph them foranalysis.


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Method-

Morphologystudy


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Method-MTT


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Method-Flowcytometry analysis

Use PI asdye.

And flow through the machineanalyzing thecontent

of DNA by theamount offluorescent

Amount of DNA

G2/M phase > Sphase(varying) > G1/G0 > apts cell

(twicetheamount of G1/G0 phase )


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Method-Flow

cytometry analysis


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Method-Flow

cytometryanalysis


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Westernbloting

Targeted protein(2nd-Ab):

Cleavagedcaspase-8,9,3,6,7

Phspho ERK ,SAPK/JNK , P-38 ERK ,SAPK/JNK , P-38

BAX

Phospho-bcl-2

Bid


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Westernbloting

Clevaged caspase-9,3,6,7


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Westernbloting


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Westernbloting


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Westernbloting


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Westernbloting


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Method-Z-VAD assay

Z-vad isa kind ofdrug that inhibit theactivity of

caspase family.

Weuse thisdrug to find out whethercaspase isreallyin thepathway of oralcavity cancercells

apoptosis.(wewant to knowsimply because the

previousexperimentsshows thepresences ofcaspase)

Thereforecellare treatedwithdifferent concentration

ofZ-VAD for I hrandat appropriate timepointsuse

MTT to determine the viability of thecells.


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!!!!^ ^


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一下 ROBERT 論文討論會台大生科 (secondary paper discussion in NTULS )

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Transcript of 一下 ROBERT 論文討論會台大生科 (secondary paper discussion in NTULS )

一下 ROBERT 論文討論會 台大生科 (secondary paper discussion in NTULS )

Documents

Transcript of 一下 ROBERT 論文討論會 台大生科 (secondary paper discussion in NTULS )

一下 ROBERT 論文討論會台大生科 (secondary paper discussion in NTULS )

Transcript of 一下 ROBERT 論文討論會台大生科 (secondary paper discussion in NTULS )