Lysophosphatidic Acid Upregulates Laminin-332 Expression during A431 Cell Colony Dispersal

Figure 4

RT-PCR analysis of mRNA expression of laminin 𝛼3, 𝛽3, and 𝛾2 genes in LPA-treated A431 cells. To confirm the results of the previously performed microarrays, RT-PCR analysis was performed as described in Section 2. Briefly, A431 cells were treated with PBS or 2 𝜇M LPA for 12 h, total RNA was extracted for each sample, and cDNA was synthesized using reverse transcriptase. Samples were analyzed on a MiQ machine using FastStart SYBR Green Master Mix. The primer sets used are shown in Table 1. Results showed that LPA treatment increased Ln-332 component, 𝛼3, 𝛽3, and 𝛾2, expression (by ~2–2.5-fold), although results were not determined to be significantly different than PBS control treatments (N = 3; P =  .11,  .08, and .13, resp.).