Long noncoding RNA ZNFX1-AS1 promotes the invasion and proliferation of gastric cancer cells by regulating LIN28 and CAPR1N1

Table 1 List of all primers used to screen gastric cancer-associated long noncoding RNAs

Name	Forward-primer	Reverse-primer
GAPDH	ACCCACTCCTCCACCTTTGAC	TGTTGCTGTAGCCAAATTCGTT
H19	TACAACCACTGCACTACCTG	TGGAATGCTTGAAGGCTGCT
CCAT1	CATTGGGAAAGGTGCCGAGA	ACGCTTAGCCATACAGAGCC
HOTAIR	GGTAGAAAAAGCAACCACGAAGC	ACATAAACCTCTGTCTGTGAGTGCC
LINC00152	CTCCAGCACCTCTACCTGTTG	GGACAAGGGATTAAGACACACA
ZNFX1-AS1	CCAGTTCCACAAGGTTAC	GCAGGTAGGCAGTTAGAA
PVT1	CTTGAGAACTGTCCTTACG	CAGATGAACCAGGTGAAC
GAS5	CACAGGCATTAGACAGAA	AGGAGCAGAACCATTAAG
SNHG12	GACTTCCGGGGTAATGACAG	GCCTTCTGCTTCCCATAGAG
TUG1	TAGCAGTTCCCCAATCCTTG	CACAAATTCCCATCATTCCC
CHET1	CCCCACAAATGAAGACACT	TTCCCAACACCCTATAAGAT
SUMP1P3	ACTGGGAATGGAGGAAGA	TGAGAAAGGATTGAGGGAAAAG
GACAT3	GGGGGCTTGTTTCTTTGTGTAG	CATTCGGCTCTGACCTCTCAC
ABHD11-AS1	GAACGGGATGAAGCCATTG	GCTGATTCTGGACCTGCTG
GACAT2	TGGATGCTTACAAAGGACTGG	CTGCAATTACGGAAAGAGCTG
uc001lsz	GACGGCACCTACTACACCTT	GCTGACCACCTTGTTGTTGAA

Table 2 The sequences of ZNFX1-AS1-shRNA1, ZNFX1-AS1-shRNA2 and NC-shRNA

	Sequence
ZFAS1-shRNA1	ACCGGTCGGGGGCCCAGGGTGGAGAGCACGAGGGCCTGGCCCAGGCACGGCCGGCGCCTCCGCCCTCGAGGAGGGCGTCACCTCAGCTCCCCCCGGCGGCGGAGCCGGCGGGCTCAGGCGGGCGCGGCTGAGGGGAGCGGACCGCGGGGGGCGGGAGATGACTGCGCCCAAGGCCTTTGCGGGCCTCAGCCGGCCCCAGAGGAAGGGGAACCCGTCGAGCGGTTTGGTGCGTGTGAAGCGCGACATGGCGAGGAAGCGGACAAGCCCGGGTGGCCCGGCGTGTAGAGGGAAGGGGGCGGGGCTAGACGCGGCCTGGACAACTACTAGAGCGCCTCGGGCTGTGCTGCTCGAGACTACATTTCCCAGAGCGACGCGCGCGGAGCGGGCGGGAAAGAGAGCGTTTCGGGTCCAGTGCGCAGGTGCGAAAGCCATCTTTGGTTATATAAGGGAGGTTCAGGAAGCCATTCGTTCTTTCGCGTCTGCGGTGCCCGGAGTGTGGTACTTCTCCTAGTTGCAGTCAGGCTTCATACGCTATTGTCCTGCCCGTTAGAGCAGCCAGCGGGTACAGAATGGATTTTGGAAGAGGGAGTCACCACTGGACCTCCAAGGAAGCCACGTGCAGACATCTACAACCTTCGATCTCCTGACGAGTTTATTGTTGGCCAAAACCAGGCTTTGATTGAACCAGGATGAATGCGGGTGTTGGAAGTAGAATATATATATACATATAAAATTGGTTGGGAGCCACGTGTACCAGTGTGTGTTGATCTTGGCTTGATTCAGTCTGCCTTGTAACAGAAACTGGCGATGGAATATGAGAGGAGCCCTCTGGAAAGAAAAGGACAGACCCTGTGCTTTCATGAAAGTGAAGATCTGGCTGAACCAGTTCCACAAGGTTACTGTATACATAGCCTGAGTTTAAAAGGCTGTGCCCACTTCAAGAATGTCATTGTTAGACTTTGAAATTTCTAACTGCCTACCTGCATAAAGAAAATAAAATCTTTTAAATCAAAAGCTAGC
ZFAS1-shRNA2	TTACAAGGCAGACTGAATCTT
NC-shRNA	TTCTCCGAACGTGTCACGT

ZFAS1: ZNFX1-AS1.

Table 3 List of primers used to detect mRNA expression of long noncoding RNA binding proteins

Name	Forward-primer (5’ to 3’)	Reverse-primer (5’to 3’)
GAPDH	ACCCACTCCTCCACCTTTGAC	TGTTGCTGTAGCCAAATTCGTT
UPF1	GGTCCCTGATAATTATGGCGATG	ACGGCATAAACCTGGGAGTG
eIF4AIII	GGCATCTACGCTTACGGTTT	CAGCCAACTCTCTTGTGGGA
IGF2BP1	AGCTCCTTTATGCAGGCTCC	GAGCCTTGAATTGGGCCTCT
FMRP	CTCAAGGCTTGGCAGGGTATG	CCGTGCCCCCTATTTCTGTA
LIN28	AGATCAAAAGGAGACAGGTGCT	AATAGCCCCCACCCATTGTG
IGF2BP2	ACCCTCTCGGGTAAAGTGGA	GTTGACAACGGCGGTTTCTG
FUS	GCAAGATGGATTCCAGGGGTG	TCCAGGAAAGTGAAAAGGGGG
ZC3H7B	TGTGCAAAGGAGGAGATCGAC	ACAGACGGAGAGTCCTTGGT
IGF2BP3	CCTGGTGAAGACTGGCTACG	CCAGCACCTCCCACTGTAAAT
CAPRIN1	CTGCTGGCTGGCTAAGTCC	GGCCGAGGGCATCGTG

Table 4 General characteristics and clinicopathological factors of gastric cancer patients and healthy control subjects

Variables	Gastric cancer (n = 15)	Healthy control (n = 15)
Gender
Male	12	12
Female	3	3
Age (yr)
Mean	61	62
Range	48-70	47-80
Tumor site1
Upper third	3	-
Middle third	4	-
Lower third	8	-
Tumor size
≥ 5 cm	6	-
< 5 cm	9	-
Pathological differentiation2
Undifferentiated or poorly differentiated	12	-
Others	3	-
TNM classification3
I + II	4	-
III + IV	11	-
Lymphatic invasion3
Positive	11	-
Negative	4	-

¹According to National Comprehensive Cancer Network Clinical Practice Guidelines in Oncology: Version I, 2016.

²According to World Health Organization Classification of Tumors of the Digestive System: 4^th edition.

³According to the American Joint Committee on Cancer Staging Manual: 7^th edition.

Table 5 General characteristics and clinicopathological factors of gastric cancer patients and healthy control subjects

Variables	Healthy control (n = 60)	Gastritis/peptic ulcer (n = 60)	GIST (n = 60)	Gastric cancer (n = 60)
Gender
Male	46	37	35	46
Female	14	23	25	14
Age (yr)
Mean	61	60	60	61
Range	37-91	37-89	30-85	37-91
Tumor site1
Upper third	-	-	-	13
Middle third	-	-	-	14
Lower third	-	-	-	33
Tumor size
≥ 5 cm	-	-	-	29
< 5 cm	-	-	-	31
Pathological differentiation2
Undifferentiated or poorly differentiated	-	-	-	45
Others	-	-	-	15
TNM classification3
I + II	-	-	-	20
III + IV	-	-	-	40
Lymphatic invasion3
Positive	-	-	-	43
Negative	-	-	-	17

¹According to National Comprehensive Cancer Network Clinical Practice Guidelines in Oncology: Version I, 2016.

²According to World Health Organization Classification of Tumors of the Digestive System: 4^th edition.

³According to the American Joint Committee on Cancer Staging Manual: 7^th edition.

GIST: Gastric stromal tumor.

Table 6 Correlation between the relative expression of long noncoding RNA HULC or ZNFX1-AS1 and clinicopathologic factors

Variables	Number of cases	P value
Gender
Male	46	0.99
Female	14
Age (yr)
≤ 61	28	0.26
> 61	32
Tumor site1
Upper third	13	0.12
Middle third	14
Lower third	33
Tumor size
≥ 5 cm	29	0.01
< 5 cm	31
Pathological differentiation2
Undifferentiated or poorly differentiated	45	0.55
Others	15
TNM classification3
I + II	20	0.02
III + IV	40
Distant metastasis3
Positive	5	0.97
Negative	55
Lymphatic invasion3
Positive	43	0.03
Negative	17

¹According to National Comprehensive Cancer Network Clinical Practice Guidelines in Oncology: Version I, 2016.

²According to World Health Organization Classification of Tumors of the Digestive System: 4^th edition.

³According to the American Joint Committee on Cancer Staging Manual: 7^th edition.