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. 2019 May 14;21(7):641–652. doi: 10.1016/j.neo.2019.04.007

Figure 2.

Figure 2

Differential Cell Growth in Response to LDHA/PDHA1 Knockdown in HNSCC Cells in vitro and Xenografic Tumor Growth in vivo. (A) Respective real-time RT-PCR and Western blot analysis for LDHA/total PDHA1 (T-PDHA1)/Phosphorylated PDHA1(S293, p-PDHA1) mRNA and protein expression in HNSCC and NHOK cells. Cell growth was assayed in LDHA- and PDHA1-silencing HNSCC cells using (B) trypan exclusion and (C) MTT assays. (D) Mitochondrial SDH activity in single unit of LDHA- and PDHA1-silencing HNSCC cells was determined by the values of MTT levels divided by corresponding cell number. Red dotted line indicated the condition that SDH activity represents compatible cell number as in control cells. (E) Seahorse metabolic analysis showed an elevated OCR in LDHA deficient FaDu cells. (F) Flow cytometry based cell cycle analysis was performed in LDHA/PDHA1 deficient HNSCC cells. LDHA loss leads to a greater percentage of G0/G1 cells (red) whereas PDHA1 deficiency results in lower G0/G1 cell proportion (green). (G) MTT assay showed suppressed cell growth in response to treatments of PDK1 inhibitor DCA in HNSCC SAS and FaDu cells. (H) In vivo HNSCC-bearing xenografic tumor growth analysis showed that LDHA loss downregulated tumor growth while PDHA1-silencing results in bigger and heavier tumor mass. shLuc vector is used as a control plasmid. R.Q.: Relative Quantification. Data are presented as mean ± SEM (N≥3). ***P < .001; **P < .01; *P < .05.