br Lung cancer cell lines

Lung cancer cell lines The greatest expression of both human and rodent FFA4 transcript has been shown to be localized to the lung [61], yet the physiological role in this tissue has been understudied, as has its involvement in lung cancers. In 2015, a two-staged genome-wide association study of 11,464 lung cancer cases and 12,206 controls from eight independent study datasets identified 30 gene variants, representing 25 independent foci, that were detected in the discovery and subsequently validated in the replication stages of the analysis. The investigators revealed that of these, a variant allele (rs12415204; NG_032670.1:g.9469C > A) of the gene encoding for FFA4 (10q23.33), representing a single nucleotide variation from C to A, had the strongest positive association with a family history of lung cancer, and also had a strong association with increased risk of lung cancer [62]. Across the entirety of datasets studied, the FFA4 allele variant had a minor allele frequency ranging from 21 to 25% and stratified analysis by age, sex, and never-smoking status revealed a potentially increased risk of lung cancer association in females, never-smokers, and those under 50 years of age with the variant FFA4 allele. These data suggest that FFA4 single-nucleotide polymorphisms may increase risk of lung cancer in innately low-risk populations. Kita and colleagues investigated the roles of FFA4 and FFA1 in rat RLCNR, mouse LL/2 and human A549 lung cancer cells, all three of which were shown to express both long-chain FFA receptor transcripts [63]. Given the expression of both FFA4 and FFA1, the study paradigm made use of GW9508 in the absence or presence of the selective FFA1 antagonist GW1100. In cell proliferation assays, no change in growth rate was found upon stimulation with GW9508 alone or in combination with GW1100 in either RLCNR or LL/2 cells. Meanwhile, cell migration was significantly stimulated upon GW9508 treatment alone, and blocked by pretreating with GW1100, suggesting FFA1 mediated effects promoting cell migration. In A549 cells, GW9508 did not induce cell proliferation but GW1100 marginally, yet significantly, inhibited it. Furthermore, GW9508 significantly reduced cell migration in these cells, which was blocked further by GW1100, suggesting that FFA4 plays a role in inhibiting proliferation and migration, while FFA1 induces these effects [63]. These results were further reinforced upon FFA1 knockdown in A549 cells, which demonstrated significantly larger GW9508-mediated inhibition of proliferation compared to their control cell line. Furthermore, investigation into the expression and activity of matrix metalloproteinase –2 (MMP-2) or MMP-9, which are known to be associated with migration and angiogenesis in cancer cells, revealed that MMP-2 was equally lowered in both FFA1-knockdown and control RGDS peptide when treated with GW9508, again suggesting a role for FFA4 as the major contributor to negative regulation of migration in A549 cells [63]. In summary, this work demonstrated that FFA1 positivity, while FFA4 negatively regulates tumor progression in lung cancer cells lines.
Pancreatic cancer cell lines On the contrary to results that show that FFA4 agonism inhibits, while FFA1 agonism stimulates, migration of A549 lung cancer cells; in human PANC-1 ductal carcinoma cells, FFA4 agonism promotes, and FFA1 inhibits, malignant properties. These cells were noted to express both long-chain FFA receptors while the hamster pancreatic duct adenocarcinoma-derived HPD1NR and HPD2NR cells expressed only FFA1 and FFA4, respectively [64]. Based on the lack of expression of both FFA receptors in the hamster cells, the authors utilized GW9508 as a selective FFA1 or FFA4 agonist in HPD1NR and HPD2NR cells, respectively. One hour of treatment with GW9508 significantly decreased cell migration in HPD1NR cells, while it significantly increased cell migration in HPD2NR cells [64]. Treatment of PANC-1 cells, which express both FFA receptors with GW9508 yielded a slight but significant decrease in migration, but a marked increase in migration upon combination of the agonist with the selective FFA1 antagonist GW1100 [64]. The effects of GW1100 alone were not shown but collectively, these results demonstrate that FFA1 inhibits, while FFA4 promotes, cell migration in pancreatic cancer cells. To confirm these observations selective knockdown of either FFA4 or FFA1 in PANC-1 cells using shRNA was employed and tumorigenic and invasive properties of these cells were measured, as were MMP-2 and MMP-9 activation. Cell proliferation assays revealed that FFA4-KD cells grew the same as control cells, while FFA1-KD cells showed significantly stunted growth after 72 h [64]. Cell migration was significantly inhibited by GW9508 treatment in FFA4-KD cells compared to control, suggesting that FFA1 is modulating agonist-stimulated inhibition of migration. Meanwhile, in FFA1-KD cells, cell migration was significantly increased, suggesting that FFA4 is modulating agonist-stimulated stimulation of migration [63]. These results were confirmed with a wound-healing assay that showed that after 20 h, FFA4-KD cells migrated to the wound-space to a lesser degree compared to control cells, while FFA1-KD cells migrated to a greater degree compared to control cells. As seen in A549 cells described above, assessment of MMP-2 and MMP-9 activity revealed no changes to MMP-9 but significant reduction of MMP-2 in FFA4-KD cells and significant increases in FFA1-KD cells, suggesting that FFA4 activity regulates increased MMP-2 function. While no effects on tumor growth were reported by this study, a single cell agar based colony formation assay revealed significantly heightened colony diameter from cells expressing FFA1-KD.