Gamma-aminobutyric acid promotes human hepatocellular carcinoma growth through overexpressed gamma-aminobutyric acid A receptor α3 subunit

Table 1 Primers for GABAA receptor subunits

Subunit	Primer	Product size (bp)
α1		902
Sense	5'-TCGTCACCAGTTTCGGACC-3'
Antisense	5'-GGTTGCTGTTGGAGCGTAA-3'
α2		722
Sense	5'-TTCACAATGGGAAGAAATCAGTAG-3'
Antisense	5'-TGCATAAGCGTTGTTCTGTATCA-3'
α3		561
Sense	5'-TCGGTCTCTCCAAGTTTGTGC-3'
Antisense	5'-TTCCGTTGTCCACCAATCTGA-3'
α4		750
Sense	5'-TGAAATTCGGGAGTTATGCCTATC-3'
Antisense	5'-GGCTGAATGGGTTTGGACTG-3'
α5		826
Sense	5'-CACCATGCGCTTGACCATCTCT-3'
Antisense	5'-GCCGAACAAGACTGGGAATA-3'
α6		764
Sense	5'-TGAGGCTTACCATCAATGCTGA-3'
Antisense	5'-GACAGGTGTTGATTGTAAGATGGG-3'
β1		603
Sense	5'-GTTCTCTATGGACTCCGAATCACA-3'
Antisense	5'-ATTGGCACTCTGGTCTTGTTTG-3'
β2		640
Sense	5'-AGCTTAAGAGAAACATTGGCTACT-3'
Antisense	5'-CGATCTATGGCATTCACATCA-3'
β3		633
Sense	5'-AGTGCTGTATGGGCTCAGAATCAC-3'
Antisense	5'-CCCGGTTGCTTTCGCTCTT-3'
γ1		262
Sense	5'-GTGTTTTGCAGCCTTGATGG-3'
Antisense	5'-TGGCAATGCGTATGTGTATCCT-3'
γ2		605
Sense	5'-AAGTCCTCCGATTGAACAGCAACA-3'
Antisense	5'-CGCTGTGACATAGGAGACCTT-3'
γ3		767
Sense	5'-ACACTCCTGCCCGCTGATT-3'
Antisense	5'-TGTCTATGTGAATACGCCCTTTCC-3'
Δ		654
Sense	5'-TCACCATCACCAGCTACCACTTCA-3'
Antisense	5'-GGGCGTAAATGTCAATGGTGTC-3'
ε		632
Sense	5'-GCAGGCGGTTTGGCTATGT-3'
Antisense	5'-CGAGTAGTTATCCAGGCGGTAG-3'
θ		465
Sense	5'-TCGAGTTCTCCTCTGCTGTG -3'
Antisense	5'-TATGCAGATCCAGGGACAA-3'
π		250
Sense	5'-CGTCGAGGTCGGCAGAAGT-3'
Antisense	5'-GCGGGCATCCAGAGTGAAG-3'

Table 2 GABAA receptor subunits mRNA expression in HepG2, Chang, L-02 and 8 sets of human HCC and adjacent nontumor tissues

Set	GABAA receptor subunits
	α1	α2	α3	α4	α5	α6	β1	β2	β3	γ1	γ2	γ3	Δ	ε	θ	π
Cell line
HepG2			+++						+				+	++	++
Chang			+++										+	++	+
L-02														++
Tumor
1			+++						+				+	++
2			++						+		+			+		+
3	+		+++										+	++		++
4									+				+	++
5			++			+			++							+
6			++						+		+		+	+
7													+	++		+
8			+						+				+	++
Non-tumor
1									+					++		+
2									++					++		+
3									+					++
4									+					+		+
5									+					++
6									++					++		++
7									+					+
8									++					+

Table 3 Cell cycle of HepG2/Si-1 and HepG2/Si-Mock (mean ± SD)

	Si-1	Si-Mock
G1/G0%	56.1 ± 3.36	50.9 ± 3.14
G2/M%	14.3 ± 1.72	14.2 ± 1.43
S%1	29.5 ± 2.26	34.5 ± 2.37

¹n = 3, P < 0.05.

Table 4 Cell cycle of HepG2 treated with GABA at serial concentrations (mean ± SD)

	0μmol/L	1μmol/L	10μmol/L	20μmol/L	40μmol/L	60μmol/L
G1/G0%	59.7 ± 3.82	58.5 ± 3.63	55.6 ± 3.46	56.5 ± 3.69	45.2 ± 3.07	57.6 ± 3.83
G2/M%	13.5 ± 1.14	18.6 ± 1.27	12.5 ± 1.15	7.6 ± 0.79	17.3 ± 1.48	12.7 ± 1.53
S%1	27.2 ± 2.19	31.3 ± 2.24	32.4 ± 2.58	35.8 ± 2.47	37.6 ± 2.64	30.4 ± 2.32

¹n = 3, P < 0.05.

Table 5 Cell cycle of HepG2/Si-Mock and HepG2/Si-1 treated with or without GABA at a concentration of 40 μmol/L (mean ± SD)

	HepG2/Si-Mock	HepG2/Si-Mock+GABA	HepG2/Si-1	HepG2/Si-1+GABA
G1/G0%	51.7 ± 3.53	49.3 ± 3.23	54.5 ± 3.37	59.4 ± 3.85
G2/M%	15.7 ± 1.75	7.2 ± 0.92	17.6 ± 1.64	15.6 ± 1.69
S%	32.4 ± 2.12	43.6 ± 2.841	27.6 ± 1.92	25.5 ± 2.06

¹HepG2/Si-Mock vs HepG2/Si-Mock + GABA, n = 3, P < 0.05.