Determination of β-glucuronidase in human colorectal carcinoma cell lines

Table 1 Conditions for measuring β-glucuronidase in culture medium

Medium (mL)	Water (mL)	Acetate buffer (mL)	pH	Substrate final	Molarity (mL)	Incubation time at 37 °C (h)	Alkalinizing reagent (mL)	Final pH	Coloremeter wavelength (nm)
0.2	0.4	0.2	4.5	0.2	0.006	18	21 + 3 mL H2O	10.2	540

¹Glycine-Duponal reagent: 15.01 g of glycine dissolved in 900 mL of H₂O and brought to pH 11.7 by addition of 50% NaOH solution. Duponal (sodium lauryl sulfate) was added to produce a final concentration of 0.2% and water was added to achieve a final volume of 1 L.

Table 2 Activity of secreted β-glucuronidase in culture medium of six colorectal carcinoma cell lines

Cell line	Differentiation degree[3]	Invasiveness[3,4]	n	β-glucuronidase activity, -x ± s
CX1	Good	Low	6	1.29 ± 0.17
CCL187	Good	Low	6	1.96 ± 0.28
CCL229	Good	High	6	3.37 ± 0.34e
CCL227	Poor	High	6	2.46 ± 0.18e
CCL228	Poor	High	6	2.73 ± 0.19e
Clone A	Poor	High	6	3.22 ± 0.38

^eP < 0.001 vs CX1, CCL187