Mutation analysis of novel human liver-related putative tumor suppressor gene in hepatocellular carcinoma

Table 1 Primer sequences for the coding region of the LPTS gene

Exon	Nucleotide sequences	PCR conditions	Product size (bp)
1	Forward: 5’-CGTGCTCGAGGAGCGAGTCG-3’
	Reverse: 5’-ACCCGGCATCTTCACCAACG-3’	52 °C SD	241
2	Forward: 5’-TCCATTGCTGATGATAATGC-3’
	Reverse: 5’-CTTCCAGTCTCCTAAGAAGG-3’	50 °C SD	230
3	Forward: 5’-TGAGAGGAATGTTCTAACTC-3’
	Reverse: 5’-AGCCAAYYAYGCAAAGACAC-3’	50 °C SD	178
4	Forward: 5’-AACTACAGGCTTACCTCTCG-3’
	Reverse: 5’-AACATATTTGCATTGAGAAC-3’	55 °C SD	196
5	Forward: 5’-CAAGACTATCCACTGTTAGG-3’
	Reverse:5’-GGACAAACACGTAGATTTCAATAAC-3’	52 °C Standard	220
6	Forward: 5’-GCTGCATAGTTCATGTCTGC-3’
	Reverse: 5’-CACAGGTGAAAATCAGACAG-3’		194
7.1	Forward: 5’-CTGCCTTTTAACTCTTCTGC-3’
	Reverse: 5’-GGCTGGAGGTAACTTTCCAC-3’	50 °C SD	247
7.2	Forward: 5’-GGCCACAGGTAAAGATGTGG-3’
	Reverse: 5’-CAGGCGGCTGCACATGGTCC-3’	55 °C Standard	200
7.3	Forward: 5’-GCCTCTGCTCAGGATGCAGG-3’
	Reverse: 5’-GCCCCGGCTGGGAAGGATTC-3’	50 °C SD	191

SD: step-down PCR

Table 2 The alterations of LPTS-S and LPTS-L in tumor cell lines

Cell line	Cell type	Mutation	Exon	Codon	Predicted effect
CCLP1	Cholangiocarcinoma	C161T	Exon 2	gct→gtt	Ala→Val
HepG2	HCC	A701A, G(LPTS-S)	Exon 7	Outside ORF	/
		A778A, G(LPTS-L)		aca→gca	Thr→Ala
HepT1	HCC	C183T	Exon 2	gcc→gct	Ala
MEK	HCC	A701A, G(LPTS-S)	Exon 7	Outside ORF	/
		A778A, G(LPTS-L)		aca→gca	Thr→Ala
SNU182	HCC	A701A, G(LPTS-S)	Exon 7	Outside ORF	/
		A778A, G(LPTS-L)		aca→gca	Thr→Ala
Li7A	HCC	A803T (LPTS-S)	Exon 7	Outside ORF	/
		A880T (LPTS-L)		agc→tgc	Ser→Cys
Li21	HCC	C183T	Exon 2	gcc→gct	Ala
		A803T (LPTS-S)	Exon 7	Outside ORF	/
		A880T (LPTS-L)		agc→tgc	Ser→Cys
MZCHA1	HCC	A803T (LPTS-S)	Exon 7	Outside ORF	/
		A880T (LPTS-L)		agc→tgc	Ser→Cys
PCI-SG231	HCC	G111C	Exon 1	Before ATG	/
		A803T (LPTS-S)	Exon 7	Outside ORF	/
		A880T (LPTS-L)		agc→tgc	Ser→Cys
PLCPRF5	HCC	A497G	Exon5	tat→tgt	Tyr→Cys
IGR-OV1	Ovarian tumor	A701A, G (LPTS-S)	Exon 7	Outside ORF	/
		A778A, G (LPTS-L)		aca→gca	Thr→Ala
SW579	Head and neck tumor	C183C, T	Exon 2	gcc→gct	Ala