Glutamine regulates the cellular proliferation and cell cycle progression by modulating the mTOR mediated protein levels of β-TrCP

The amino acid glutamine plays an important role in cell growth and proliferation. Reliance on glutamine has long been considered a hallmark of highly proliferating cancer cells. Development of strategies for cancer therapy that primarily target glutamine metabolism has been an active area of research. Glutamine depletion is associated with growth arrest and apoptosis-induced cell death; however, the molecular mechanisms involved in this process are not clearly understood. Here, we show that glutamine depletion activates the energetic stress AMPK pathway and inhibits mTORC1 activity. Furthermore, inhibition of mTORC1 reduces the protein levels of β-TrCP, resulting in aberrant cell cycle progression and reduced proliferation. In agreement with the role of β-TrCP in glutamine metabolism, knockdown of β-TrCP resulted in proliferation and cell cycle defects similar to those observed for glutamine depletion. In summary, our results provide mechanistic insights into the role of glutamine metabolism in regulation of cell growth and proliferation via β-TrCP, uncovering a previously undescribed molecular process involved in glutamine metabolism.</p

Supplementary figures 1-9 Supplementary Table 1 from Long Noncoding RNA MALAT1 Regulates Cancer Glucose Metabolism by Enhancing mTOR-Mediated Translation of TCF7L2

Supplementary Information: Figure S1: MALAT1 affects cancer glucose metabolism Figure S2: Glucose metabolism in HCC cell lines with MALAT1 knockdown. Figure S3: Regulation of TCF7L2 protein expression by MALAT1 in HCC cell lines. Figure S4: A non-phosphorylatable mutant of 4EBP1 inhibits TCF7L2 protein expression and expression of glycolytic genes. Figure S5: SRSF1 regulates TCF7L2 levels post-transcriptionally. Figure S6: TCF7L2 modulates glucose metabolism in a HCC cell line. Figure S7: MALAT1 and TCF7L2 regulate gluconeogenesis through the same pathway. Figure S8: Oncogenic properties of HCC cell lines with TCF7L2 knockdown. Figure S9: TCF7L2 protein, Gluconeogenesis and Glycolytic enzyme expression in livers from mouse HCC model Mdr2-/-. Table S1: List and sequences of shRNAs, siRNAs and PCR primers used in the paper</p

Table S1 from Long Noncoding RNA MALAT1 Promotes Hepatocellular Carcinoma Development by SRSF1 Upregulation and mTOR Activation

Table S1: List of primers oligos and siRNAs</p

Table S3 from Long Noncoding RNA MALAT1 Promotes Hepatocellular Carcinoma Development by SRSF1 Upregulation and mTOR Activation

Table S3. IPA enriched pathways based on RNA-seq analysis</p

Figures S1-S8 from Long Noncoding RNA MALAT1 Promotes Hepatocellular Carcinoma Development by SRSF1 Upregulation and mTOR Activation

Figure S1: Knockdown of MALAT1 inhibits proliferation of liver progenitor and HCC cells. Figure S2: Effect of MALAT1 expression on splicing of endogenous SRSF1 targets. Figure S3. Differential gene expression based on RNA-seq data. Figure S4. Enriched pathways and networks activated by overexpression of MALAT1 based on RNA-seq analysis. Figure S5 Enriched pathways activated by MALAT1 overexpression based on RNA-seq analysis. Figure S6. Validation of MALAT1 up- and down-regulated genes identified by RNA-seq analysis. Figure S7. Knockdown of MALAT1 down-regulates c-Myc protein levels. Figure S8. Knockdown of SRSF1 inhibits oncogenesis downstream to MALAT1 and only partially inhibits transformation by oncogenic Ras.</p

Table S2 from Long Noncoding RNA MALAT1 Promotes Hepatocellular Carcinoma Development by SRSF1 Upregulation and mTOR Activation

Table S2. Full list of Up- and Down regulated genes in the RNA seq analysis</p

Supplementary Materials and Methods and Supplementary Figure Legends from Long Noncoding RNA MALAT1 Promotes Hepatocellular Carcinoma Development by SRSF1 Upregulation and mTOR Activation

Supplementary information text: Methods and figure legends</p