The effects of REG4 expression on chemoresistance of ovarian cancer

Figures & data

Table 1. Primary antibodies used in the present study.

Name	Dilution	Source	Company
GAPDH	1:2000	Rabbit	CST,USA
REG4	1:1000	Goat	R&D, USA
Pi3-Kinase p85/p55	1:1000	Rabbit	Immunoway, USA
Pi3K	1:1000	Rabbit	Immunoway, USA
P-AKT	1:1000	Rabbit	CST,USA
AKT	1:1000	Rabbit	CST,USA
P-mTOR	1:1000	Rabbit	CST,USA
mTOR	1:1000	Rabbit	CST,USA
Bcl2	1:1000	Rabbit	CST,USA
Survivin	1:500	Mouse	Santa Cruz, USA
GST-π	1:1000	Rabbit	Abcam, USA

Figure 1. Expression of REG4 mRNA in ovarian cancer. (A) Regenerating gene 4 (REG4) mRNA expression was observed in various cancers according to an Oncomine database. (B) A positive correlation existed between REG4 mRNA expression and cell purity in ovarian cancer according to a Timer database. (C) The prognostic significance of REG4 mRNA expression was analysed in ovarian cancer treated with taxol chemotherapy using a Kaplan–Meier plotter. (D) The prognostic significance of REG4 mRNA expression was analysed in ovarian cancer treated with platinum (platin) chemotherapy using a Kaplan–Meier plotter.

Table 2. Prognostic value of REG4 in subtype ovarian cancer treated with platin.

Clinicopathological features	Overall survival		Progression-free survival		Post-progression survival
	Hazard ratio	p	Hazard ratio	p	Hazard ratio	p
Histology
Endometrioid	–	–	–	–	–	–
Serous	1.57 (1.22–2.02)	<.001	1.56 (1.24–1.95)	<.001	1.27 (0.98–1.64)	.070
Clinical Stage
I + II	0.32 (0.09–1.17)	.070	0.32 (0.13–0.76)	.007	–	–
III + IV	1.61 (1.27–2.05)	<.001	1.56 (1.25–1.93)	<.001	1.34 (1.05–1.71)	.019
Grade
I + II	1.59 (1.01–2.51)	.043	0.75 (0.52–1.08)	.116	1.66 (1.04–2.65)	.032
III	1.59 (1.18–2.14)	.002	1.65 (1.26–2.17)	<.001	1.25 (0.91–1.73)	.167
TP53 mutation
Mutated	1.39 (0.93–2.06)	.107	1.92 (1.31–2.81)	<.001	0.75 (0.49–1.14)	.174
Wild type	2.26 (0.71–7.26)	.159	2.13 (0.73–6.22)	.156	1.65 (0.51–5.34)	.400

Table 3. Prognostic value of REG4 in ovarian cancer treated with paclitaxel.

Clinicopathological features	Overall survival		Progression-free survival		Post-progression survival
	Hazard ratio	p	Hazard ratio	p	Hazard ratio	p
Histology
Endometrioid	–	–	–	–	–	–
Serous	1.60 (1.17–2.18)	.003	1.43 (1.1–1.87)	.008	1.28 (0.93–1.76)	.124
Clinical stage
I + II	3.77 (0.32–44.33)	.263	0.26 (0.08–0.79)	.010	–	–
III + IV	1.66 (1.23–2.23)	<.001	1.47 (1.13–1.90)	.004	1.40 (1.05–1.88)	.023
Pathological grade
I + II	1.70 (0.98–2.93)	.055	0.75 (0.50–1.13)	.164	1.53 (0.88–2.67)	.129
III	1.48 (1.02–2.15)	.037	1.60 (1.15–2.22)	.005	1.26 (0.86–1.85)	.240
TP53 mutation
Mutated	1.67 (0.97–2.86)	.062	2.07 (1.2–3.56)	.008	1.54 (0.9–2.64)	.115
Wild type	–	–	–	–	–	–

Figure 2. Effects of REG4 on cellular phenotypes of ovarian cancer cells. (A) After transfection of a regenerating gene 4 (REG4)-overexpressing plasmid, REG4 mRNA was detected in CAOV3 cells by real-time reverse transcriptase (RT)–PCR. (B) After transfection of REG4-overexpressing plasmid, REG4 protein was detected in CAOV3 cells by western blot. (C) The effect of REG4 expression on the proliferation of CAOV3 cells treated with cisplatin or taxol. (D) The effect of REG4 expression on apoptosis in CAOV3 cells treated with cisplatin or taxol. (E) The effect of REG4 expression on migration and invasion by CAOV3 cells treated with cisplatin or taxol. Note: compared with control, *p < .05; **p < .01; ***p < .001.

Figure 3. Effect of REG4 expression on expression of drug-resistant proteins in ovarian cancer cells. Western blotting was used to screen chemoresistance-related proteins in ovarian cancer cells or their regenerating gene 4 (REG4) transfectants treated with cisplatin or taxol. Note: REG4-transfected cells were compared with mock-transfected cells, *p < .05; **p < .01; compared with cisplatin or taxol, ^#p < .05; ^##p < .01.

Data availability statement

There is no data needed to be deposited. The datasets generated and/or analysed during the current study are available from the corresponding author on reasonable request. This study was performed according to the Enhancing the QUAlity and Transparentcy of health Research (EQUATOR) network guideline.