Protein microarray analysis of apoptosis-related protein expression following heat shock in human tongue squamous cell carcinomas containing different p53 phenotypes

Figures & data

Table I.  Survival and apoptosis rates in SAS/neo and SAS/mp53 cells after heating (44°C, 40 min) [15].

	(%)
Cells	Survival^1	Apoptosis^2
SAS/neo	9.0	32.0 ± 6.6*
SAS/mp53	23.3	4.8 ± 3.5*

¹ Survival rate was determined using colony formation assays. ² Apoptosis was detected with Hoechst 33342 staining at 48 h after heating (44°C, 40 min). Observations were made with a fluorescence microscope. The staining analysis was performed at three replicate points per point. (Mean ± standard errors). *p < 0.05.

Figure 1. Unsupervised hierarchical clustering analysis of 512 proteins in heat treated cells compared with non-treated cells. The graph indicates relative protein expression levels. Lower signal values are indicated by green, and higher signal values are indicated by red.

Figure 2. Hierarchical clustering analysis of 44 selected apoptosis-related proteins which showed valid changes in heat treated cells when compared to non-treated cells. The graph indicates protein expression levels with lower signal values shown in green and higher signal values shown in red.

Figure 3. INRs for apoptosis-related proteins which were obtained from protein microarray analysis. The central line is the average value for INRs as measured from the expression of 44 selected apoptosis-related proteins which showed valid changes in response to heat treatment.

Table II.  Apoptosis-related proteins with more than a 1.5-fold increase in expression in SAS/neo cells.

Apoptosis	Proteins	INR values
Promoters of apoptosis	FADD/Mort-1	1.53
Mediators of apoptosis	Ku70	1.86
	E2F-1	1.62
	PKCε	1.59
	Hsp60	1.52
	PKCδ	1.52
Inhibitors of apoptosis	PARP	1.59

Table III.  Apoptosis-related proteins with more than a 1.5-fold increase in expression in SAS/mp53 cells.

Apoptosis	Proteins	INR values
Promoters of apoptosis	Fos	4.41
	Baxα	2.47
	Smad4/DPC4	2.45
	cRAF-1	2.44
	Smad2	2.42
	Bak	2.36
	TGF-β1	2.27
	c-Myc	2.21
	FADD/Mort-1	2.14
	DAP3	1.59
Mediators of apoptosis	PKCδ	2.64
	Cytochrome c/Apaf-2	2.55
	MST1	2.48
	Cdk2	2.40
	Ku70	2.36
	p21	2.35
	Hsp60	2.35
	E2F-1	2.32
	Ras (Ha)	2.31
	PKCε	2.29
	Caspase-14	2.26
	Ku80	2.24
	TRAF4	2.24
	p38d/SAPK4	2.24
	PI-3 Kinase (p110α)	2.19
	ERK1	1.97
Inhibitors of apoptosis	STAT3	2.93
	DFF45 (ICAD)	2.73
	NF-κB	2.61
	PKCλ	2.61
	PKCθ	2.60
	Bcl-2	2.44
	PKCβ	2.41
	PKCα	2.35
	Rb2	2.34
	COX2/PGHS	2.34
	IKKα/1	2.25
	MDM2	2.25
	Hsp90	2.22
	Bcl-xL	2.20
	PKCι	2.14
	IL-6	2.11
	PARP	2.00
	TRAF2	1.69

Figure 4. A proposed model for a heat-induced signal transduction pathway based on data obtained from protein microarray analysis.

Yasumoto J, Kirita T, Takahashi A, Ohnishi K, Imai Y, Yuki K, Ohnishi T. Apoptosis-related gene expression after hyperthermia in human tongue squamous cell carcinoma cells harboring wild-type or mutated-type p53. Cancer Lett 2004; 204: 41–51

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