Impact of high-mobility-group A2 overexpression on epithelial-mesenchymal transition in pancreatic cancer

Figures & data

Figure 1 Immunohistochemical staining of HMGA2, E-cadherin and vimentin in normal pancreatic tissue and pancreatic cancer samples. (A) No expression of HMGA2 in the normal pancreatic tissue. (B) Positive signals of HMGA2 in pancreatic cancer. (C) No expression of vimentin in the normal pancreatic tissue. (D) Positive signals of vimentin in pancreatic cancer. (E) Positive signals of E-cadherin in the normal pancreatic tissue. (F): No expression of E-cadherin in pancreatic cancer. Scale bar=50 um.

Table 1 Association of HMGA2, E-cardherin and Vimentin expression with the clinicopathological features of 60 patients with pancreatic cancer

Characteristics	Cases	HMGA2 expression			E-cardherin expression			Vimentin expression
Positive	Negative	P	Positive	Negative	P	Positive	Negative	P
Age				0.7563			0.6753			0.805
≤60	26	11	15		12	14		10	16
61–70	16	6	10		6	10		9	7
>70	18	9	9		6	12		8	10
Gender				0.2981			1			0.4313
Male	37	14	23		15	22		15	22
Female	23	12	11		9	14		12	11
Treatment				1			0.2952			0.1932
No treatment	33	15	18		11	22		12	21
Gemcitabine	27	12	15		13	14		15	12
Clinical stage				0.0285			0.0344			0.0051
I	11	1	10		8	3		2	9
Ⅱ	43	21	22		15	28		19	24
III/IV	6	4	2		1	5		6	0
Tumor Differentiation				0.0419			0.0301			0.009
Well differentiated	8	1	7		6	2		2	6
Mediated differentiated	33	13	20		14	19		11	22
Poor differentiated	19	12	7		4	15		14	5
Lymph node metastasis				0.0006			0.0326			0.0086
Yes	26	18	8		6	20		17	9
No	34	8	26		18	16		10	24

Note: Statistically significant values are shown in bold. 

Figure 2 Survival curves of patients with HMGA2, E-cadherin and vimentin expression in pancreatic cancer. (A) Patients with high HMGA2 levels showed significantly worse survival than those with low expression (P=0.022). (B) Patients with high vimentin levels showed significantly worse survival than those with low expression (P=0.028). (C) Patients with low E-cadherin levels had significantly worse survival than those with high expression (P=0.003).

Table 2 Univariate and multivariate analysis of factors associated with overall- survival of patients with pancreatic cancer

	Cases	Univariable			Multivariable
HR	95%CI	P	HR	95%CI	P
Age
≤60	26	1
>60	34	0.713	0.212–1.257	0.383
Gender
Male	37	1
Female	23	0.461	0.292–1.321	0.421
Treatment
No treatment	33	1			1
Gemcitabine	27	0.342	0.127–0.764	0.012	0.669	0.412–0.865	0.032
Clinical stage
I	11	1			1
II/III/IV	49	4.239	2.126–8.432	0.003	8.521	1.743–32.129	0.026
Tumor Differentiation
Well/differentiated	41	1			1
Poor differentiated	19	4.569	1.539–9.439	0.011	1.643	0.632–2.823	0.727
Lymph node metastasis
No	34	1			1
Yes	26	3.153	1.356–6.539	0.001	6.714	1.867–13.524	0.015
HMGA2 expression
Negative	34	1			1
Positive	26	3.216	1.238–6.427	0.001	5.712	1.128–13.572	0.038
E-cadherin expression
Negative	36	1			1
Positive	24	0.394	0.164–0.715	0.013	0.846	0.226–4.921	0.561
Vimentin expression
Negative	33	1			1
Positive	27	4.936	1.133–8.316	0.004	0.964	0.493–3.427	0.096

Note: Statistically significant values are shown in bold. 

Figure 3 HMGA2 knockdown in pancreatic cancer CAPAN 1 cells using SiRNAs (HMGA2-S1 and HMGA2-S2). (A) Quantitative real-time PCR and (B) Western blot data showing decreased HMGA2 mRNA and protein expression levels in both knockdown groups. ***P<0.001. HMGA2 over expressed in PC cells using plasmid pcDNA HMGA2 and non-target controls (pcDNA). (C) Quantitative real-time PCR and (D) Western blot data showing increased HMGA2 mRNA and protein expression levels in pcDNA HMGA2 group. **P<0.01.

Figure 4 (A) Compared with control cells, migration and invasion abilities in HMGA2-S1 and HMGA2-S2 cells were significantly inhibited (x100). (B) Columns show the means of three independent experiments. *P<0.05, ***P<0.001. (C) Compared with control cells, migration and invasion abilities in HMGA2 overexpressed cells were significantly enhanced (x100). (B) Columns show the means of three independent experiments. *P<0.05, **P<0.01.

Figure 5 (A) Western blot for detecting EMT-associated protein markers in HMGA2 knockdown and overexpressed CAPAN 1 cells. (B) and (C) Semiquantitative histograms of Western blot results. *P<0.05, **P<0.01, ***P<0.001.