Validation and analysis of key factors of Banxia Xiexin decoction against gastric cancer

Table 1 Sequence list of PCR primers

Primer name	Upstream primer sequence (5’-3’)	Downstream primer sequences (5’-3’)
AKT1	GCTCAGCCCACCCTTCAAG	GCTGTCATCTTGGTCAGGTGGT
PIK3CA	TAGTGTCCGGGAAAATGGCT	GGCATGCTCTTCGATCACAG
TP53	GCTGCTCAGATAGCGATGGTCT	CAACCTCAGGCGGCTCATAG
HSP90AA1	GAAGGAATTTGAGGGGAAGACTTTA	TGCCATGTAACCCATTGTTGAG
PIK3R1	AGCAACCTGGCAGAATTACG	GCTGCTGGAATGACAGGATT
GAPDH	GAAGGTCGGAGTCAACGGAT	CCTGGAAGATGGTGATGGG

AKT1: AKT serine/threonine kinase 1; PIK3CA: Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha; TP53: Tumor protein p53; HSP90AA1: Heat shock protein 90 alpha family class A member 1; PIK3R1: Phosphoinositide-3-kinase regulatory subunit 1; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Table 2 IC₅₀ screening and cell proliferation assay of Cell Counting Kit-8 for the detection of Banxia Xiexin decoction in AGS cells

Group	Volume fraction, %	24 h	24-h cell viability, %	48 h	48-h cell viability, %
AGS	0	2.050 ± 0.044	100	2.469 ± 0.004	100
BXD	10	1.628 ± 0.024b	78.48	1.983 ± 0.023b	79.57
	20	1.316 ± 0.010b	62.54	1.382 ± 0.064b	54.30
	40	0.661 ± 0.008b	29.13	0.529 ± 0.027b	18.46

^bP < 0.05. BXD: Banxia Xiexin decoction.

Table 3 Comparison of relative expression of phosphatidylinositol 3-kinase/AKT signaling pathway related proteins in each group

Group	PIK3CA	HSP90AA1	PIK3R1	AKT1	TP53
GES-1	0.27 ± 0.007	0.29 ± 0.05	0.22 ± 0.002	0.21 ± 0.005	0.24 ± 0.03
AGS	0.82 ± 0.036b	0.96 ± 0.029b	0.87 ± 0.014b	0.9 ± 0.085a	0.85 ± 0.032b
Capecitabine	0.45 ± 0.05c	0.57 ± 0.018d	0.54 ± 0.01d	0.48 ± 0.03c	0.55 ± 0.033c
BXD	0.59 ± 0.05	0.72 ± 0.04c	0.73 ± 0.021c	0.63 ± 0.007	0.63 ± 0.063
LY294002	0.57 ± 0.015c	0.64 ± 0.039c	0.66 ± 0.005d	0.57 ± 0.013	0.61 ± 0.006c
BXD + LY	0.37 ± 0.023d	0.42 ± 0.006d	0.47 ± 0.009d	0.34 ± 0.022c	0.37 ± 0.028d

^aP < 0.05 vs GES-1.

^bP < 0.01 vs GES-1.

^cP < 0.05 vs AGS.

^dP < 0.01 vs AGS.

PIK3CA: Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha; HSP90AA1: Heat shock protein 90 alpha family class A member 1; PIK3R1: Phosphoinositide-3-kinase regulatory subunit 1; AKT1: AKT serine/threonine kinase 1; TP53: Tumor protein p53; BXD: Banxia Xiexin decoction; LY: LY294002.

Table 4 Comparison of relative expression of phosphatidylinositol 3-kinase/AKT signaling pathway mRNA in each group

Group	PIK3CA	HSP90AA1	PIK3R1	AKT1	TP53
GES-1	1.00 ± 0.00	1.00 ± 0.00	1.00 ± 0.00	1.00 ± 0.00	1.00 ± 0.00
AGS	1.97 ± 0.011b	3.73 ± 0.86a	5.75 ± 0.77b	1.86 ± 0.09b	4.66 ± 2.11a
Capecitabine	0.44 ± 0.05d	1.26 ± 0.11	2.81 ± 1.41	0.47 ± 015d	0.13 ± 0.01c
BXD	0.85 ± 0.09c	1.34 ± 0.19	0.55 ± 0.31c	0.91 ± 0.20d	0.21 ± 0.09c
LY294002	1.61 ± 0.22	2.01 ± 0.55	0.52 ± 0.08d	0.84 ± 0.03d	0.27 ± 0.16c
BXD + LY	0.14 ± 0.11d	1.11 ± 0.05	0.46 ± 0.35d	0.88 ± 0.17d	0.17 ± 0.02c

^aP < 0.05 vs GES-1.

^bP < 0.01 vs GES-1.

^cP < 0.05 vs AGS.

^dP < 0.01 vs AGS.

PIK3CA: Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha; HSP90AA1: Heat shock protein 90 alpha family class A member 1; PIK3R1: Phosphoinositide-3-kinase regulatory subunit 1; AKT1: AKT serine/threonine kinase 1; TP53: Tumor protein p53; BXD: Banxia Xiexin decoction; LY: LY294002.