The expression of aplysia ras homolog I (ARHI) and its inhibitory effect on cell biological behavior in esophageal squamous cell carcinoma

Figures & data

Figure 1 Immunostaining demonstrating the expression of ARHI in ESCC tissues.

Notes: (A) High expression of ARHI in ESCC, ×100. (B) High expression of ARHI in ESCC, ×200. (C) Low expression of ARHI in ESCC, ×100. (D) Low expression of ARHI in ESCC, ×200.
Abbreviations: ARHI, aplysia ras homolog I; ESCC, esophageal squamous cell carcinoma.

Table 1 Relationship between ARHI expression and clinical pathological features

Values	n	ARHI		χ^2	P-value
		High	Low
Gender				1.109	0.297
Female	4	2	2
Male	77	20	57
Age (years)				0.088	0.807
≤60	39	10	29
>60	42	12	30
Tumor invasion				0.657	0.426
T1–2	57	14	43
T3–4	24	8	16
Lymph nodes metastasis				8.437	0.004*
N0	45	18	27
N1–3	36	4	32
Tumor metastasis				0.541	0.472
M0	79	21	58
M1	2	1	1
TNM stage				3.126	0.086
I–II	46	16	30
III–IV	35	6	29
Differentiation				5.059	0.08
Good	26	11	15
Moderate	45	10	35
Poor	10	1	9

Note:

* Statistical significance at P<0.05.

Abbreviations: ARHI, aplysia ras homolog I; TNM, tumor node metastasis.

Figure 2 (A) Kaplan–Meier analysis of the correlation between ARHI expression level and overall survival; (B) the relative expression levels of ARHI were determined by western blotting; (C) cell proliferation was assessed daily for 5 days using the Cell Counting Kit-8 assay in ESCC cells.

Abbreviations: ARHI, aplysia ras homolog I; ESCC, esophageal squamous cell carcinoma.

Table 2 Univariate and multivariate analyses of survival in ESCC

Variable	n	Univariate		Multivariate
		HR (95% CI)	P-value	HR (95% CI)	P-value
Age (years)
≤60	39	1
>60	42	1.473 (0.877–2.472)	0.143
Sex
Male	77	1
Female	4	0.244 (0.034–1.769)	0.163
T stage
pT1–pT2	57	1
pT3–pT4	24	0.981 (0.391–2.459)	0.967
N stage
pN0	45	1		1
pN1–pN2	36	3.196 (1.871–5.460)	<0.001	2.803 (1.626–4.831)	<0.001*
M stage
pM0	79	1
pM1	2	3.101 (0.742–12.971)	0.121
TNM stage
I–II	46	1
III–IV	35	2.477 (1.464–4.192)	0.001*
Differentiation
Good	26	1
Moderate-poor	55	1.475 (0.971–2.239)	0.068
ARHI expression
Low	59	1		1
High	22	0.370 (0.186–0.738)	0.005*	0.458 (0.227–0.924)	0.029*

Note:

* Statistical significance at P<0.05.

Abbreviations: ARHI, aplysia ras homolog I; HR, hazard ratio; CI, confidence interval; TNM, tumor node metastasis.

Figure 3 (A) Transwell assays were used to assay the involvement of ARHI for invasion in ESCC cells, original magnification ×200; (B) the cell counts of the ECA109 group, control group, and ARHI group; (C) wound assays were monitored at 0, 24, 48, and 72 h in ESCC cells, original magnification ×100. **P<0.01.

Abbreviations: ARHI, aplysia ras homolog I; ESCC, esophageal squamous cell carcinoma; HPF, high power field.

Figure 4 (A) The translational levels of invasive, autophagy, and EMT-related markers were determined by western blotting; (B) cells were stained with both Annexin V-APC and PI, and cells that were positive for Annexin V-APC were counted as apoptotic cells; (C) the proportion of apoptotic cells for ECA109 group, control group, and ARHI group, **P<0.01.

Abbreviations: ARHI, aplysia ras homolog I; EMT, epithelial–mesenchymal transition; APC, allophycocyanin; PI, propidium iodide.

Figure S1 (A) Cell proliferation was assessed daily for 5 days using the Cell Counting Kit-8 assay in ECA109 group, Control-siRNA group and ARHI-siRNA group; (B) Transwell assays were used to examine the involvement of ARHI for invasion in ECA109 group, control–siRNA group, and ARHI–siRNA group (original magnification ×200); (C) the cell counts of the ECA109 group, control–siRNA group, and ARHI–siRNA group in transwell assays.

Notes: *P<0.05, **P<0.01.

Abbreviations: ARHI, aplysia ras homolog I; siRNA, small interfering RNA; HPF, high power field.