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. 2021 Apr 6;12(4):368. doi: 10.1038/s41419-021-03645-4

Fig. 7. IL-6 derived from CAFs promote MALAT1 expression in GC cells.

Fig. 7

A Relative expression of MALAT1 was significantly higher in MKN-45 or MGC-803 co-cultured with CAFs than those co-cultured with NFs (CAF/MKN-45vs NF/MKN-45: 1.51 ± 0.13 vs 0.99 ± 0.01; CAF/MGC-803 vs NF/MGC-803: 1.95 ± 0.23 vs 0.99 ± 0.01, *P < 0.05, **P < 0.01); B GESA dataset analysis showed that IL-6/STAT3 pathway signalling had a positive association with MALAT1 expression(NES = 1.459, FDR q-value=0.26); C IL-6 was highly expressed in CAFs compared to NFs and GC cells(*P < 0.05, **P < 0.01); D The effect of reIL-6 (100 ng/mL) on MALAT1 expression in MKN-45 and MGC-803 were measured by qRT-PCR (MKN-45/IL-6 vs MKN-45/PBS: 1.86 ± 0.27 vs 1.03 ± 0.01: MGC-803/IL-6 vs MGC-803/PBS: 2.15 ± 0.03 vs 1.03 ± 0.01, *P < 0.05, **P < 0.01); E The effect of CAFs on MALAT1 expression in MKN-45 and MGC-803 cells was determined with the presence of IL-6 neutralizing antibody or IgG isotype control antibody (CAF + Anti-IgG/MKN-45 vs CAF + Anti-IL-6/MKN-45: 1.38 ± 0.04 vs. 1.02 ± 0.03; CAF + Anti-IgG/MGC-803 vs CAF + Anti-IL-6/MGC-803: 1.51 ± 0.04 vs 1.02 ± 0.03, **P < 0.01); F, G GEO dataset analysis suggested MALAT1 expression was positively associated with IL-6 and STAT3 expressions, respectively; H STAT3 upregulation promoted MALAT1 expression in MKN-45 and MGC-803 cells (STAT3/MKN-45 vs NC/MKN-45: 2.16 ± 0.16 vs 1: STAT3/MGC-803 vs NC/MGC-803: 3.46 ± 0.27 vs 1.1 ± 0.01, P < 0.01); I WP1066, a selective STAT3 inhibitor, could attenuate MALAT1 expression induced by recombinant IL-6 protein, which was measured by qRT-PCR (IL-6 + WP1066/MKN-45 vs IL-6+DMSO/MKN-45: 0.60 ± 0.07 vs 0.99 ± 0.03: IL-6 + WP1066/MGC-803 vs IL-6+DMSO/MGC-803: 0.24 ± 0.01 vs 0.99 ± 0.03, P < 0.01); J Potential STAT3 bindings sites on MALAT1 promoter; K Luciferase activity was measured after transfecting with MALAT1 promoter truncations, indicating that MALAT1 promoter site#3 contains binding sites (STAT3 vs NC: 3.91 ± 0.33 Vs 1.13 ± 0.17, P < 0.01); L Chip assay was performed to show that STAT3 could physically bind to MALAT1 promoter site#3 in MKN-45 and MGC-803 cells. Bars, S.D.; *P < 0.05; **P < 0.01; ***P < 0.001.