Expression of N-Acetylgalactosamine 4-Sulfate 6-O-Sulfotransferase Involved in Chondroitin Sulfate Synthesis Is Responsible for Pulmonary Metastasis

Table 1

Disaccharide composition of CS chains in the control- and 4S6ST-shRNA/LLC cells. The GAG-peptide preparation from each cell line was digested with a mixture of chondroitinases ABC and AC-II and analyzed by anion-exchange HPLC after labeling with a fluorophore 2AB as detailed in Section 2.

	Wild type	GalNAc4S-6ST-shRNA			Control-shRNA
		No. 7	No. 17	No. 23	No. 5	No. 10	No. 14
	pmol/mg acetone powder (mol%)^b
ΔA^a	675 (93.6)	209 (98.1)	321 (99.4)	243 (98.9)	1,273 (94.4)	867 (93.8)	487 (91.0)
ΔE^a	46 (6.4)	4 (1.9)	2 (0.5)	3 (1.1)	75 (5.6)	57 (6.2)	48 (9.0)
Total	721 (100)	213 (100)	323 (100)	246 (100)	1,348 (100)	924 (100)	535 (100)

^aΔA and ΔE represent ΔHexUA-GalNAc(4-𝑂-sulfate), and ΔHexUA-GalNAc(4-𝑂-, 6-𝑂-disulfate), respectively. No other disaccharide units including ΔO,ΔC, or ΔD were detected (data not shown). ΔO,ΔC, and ΔD stand for ΔHexUA-GalNAc, ΔHexUA-GalNAc(6-𝑂-sulfate) and ΔHexUA(2-𝑂-sulfate)-GalNAc(6-𝑂-disulfate), respectively.
bValues are expressed in pmol of disaccharide per mg acetone powder as starting materials from the cells and calculated based on the peak areas of the disaccharides detected by anion-exchange HPLC (Figures 1(b) and 1(c)).