Figure 3
Figure 3. Exogenous ATX controls LPA-dependent cell proliferation and endothelial cell transmigration. (A) Real-time PCR analysis of ATX mRNA expression in osteosarcoma, breast, and prostate cancer cell lines. Values were normalized to housekeeping L32 gene. Data represent the mean (± SD) of 2 independent experiments performed in triplicate. (B) Real-time PCR analysis of LPA receptor (LPA1-6) mRNA expression in MDA-231 and MDA-B02 cell lines. Values were normalized to housekeeping L32 gene. Data represent the mean (± SD) of 2 independent experiments performed in triplicate. (C) Cell proliferation assessed by 5-bromo-2′-deoxyuridine incorporation of MDA-MB-231 (left) and MDA-BO2 (right) cells in response to LPC and recombinant ATX in the presence or absence of BMP22. Results are expressed as the mean (± SD) of 3 independent experiments performed in 6 replicates (*P < .05; **P < .01; ***P < .001; vs cancer cells in presence of ATX and LPC using 1-way ANOVA with a Bonferroni posttest). (D) Effects of BMP22 on MDA-MB 231 cell invasion. Dose-response curves were generated in the presence of ATX and increasing concentrations of BMP22. Data represent the mean percentage of inhibition (± SEM) of 3 independent experiments performed in quadruplicate. (E) Effects of BMP22 on MDA-MB 231 cell transmigration across a HUVEC monolayer. Data represent the mean (± SEM) of 2 experiments performed in quadruplicate (***P < .0001 vs MDA-231 cells in presence of ATX and LPC using 1-way ANOVA with a Bonferroni posttest).

Exogenous ATX controls LPA-dependent cell proliferation and endothelial cell transmigration. (A) Real-time PCR analysis of ATX mRNA expression in osteosarcoma, breast, and prostate cancer cell lines. Values were normalized to housekeeping L32 gene. Data represent the mean (± SD) of 2 independent experiments performed in triplicate. (B) Real-time PCR analysis of LPA receptor (LPA1-6) mRNA expression in MDA-231 and MDA-B02 cell lines. Values were normalized to housekeeping L32 gene. Data represent the mean (± SD) of 2 independent experiments performed in triplicate. (C) Cell proliferation assessed by 5-bromo-2′-deoxyuridine incorporation of MDA-MB-231 (left) and MDA-BO2 (right) cells in response to LPC and recombinant ATX in the presence or absence of BMP22. Results are expressed as the mean (± SD) of 3 independent experiments performed in 6 replicates (*P < .05; **P < .01; ***P < .001; vs cancer cells in presence of ATX and LPC using 1-way ANOVA with a Bonferroni posttest). (D) Effects of BMP22 on MDA-MB 231 cell invasion. Dose-response curves were generated in the presence of ATX and increasing concentrations of BMP22. Data represent the mean percentage of inhibition (± SEM) of 3 independent experiments performed in quadruplicate. (E) Effects of BMP22 on MDA-MB 231 cell transmigration across a HUVEC monolayer. Data represent the mean (± SEM) of 2 experiments performed in quadruplicate (***P < .0001 vs MDA-231 cells in presence of ATX and LPC using 1-way ANOVA with a Bonferroni posttest).

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