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Role of exosomes in hepatocellular carcinoma cell mobility alteration.

ABSTRACT: Exosomes have gained increased research focus due to their key roles as messengers. The components of exosomes include proteins and RNAs that may be horizontally transferred between adjacent or distant cells. Hepatocellular carcinoma (HCC) is among the most malignant types of cancer worldwide, with exosomes implicated to play a crucial role in its regulation; however, the possible function of exosomes in modulating the motile ability of tumor cells and key molecules in HCC remain largely unknown. To investigate the regulatory effect of exosomes on the motile ability of HCC cells, exosomes from the culture medium of different HCC origins (high metastatic MHCC97-H and low metastatic MHCC97-L cells) were isolated for in vitro migration and invasion assays. The results indicated that the motile ability of MHCC97-L cells was significantly increased by pretreatment with MHCC97-H-derived exosomes when compared with MHCC97-L-exosome pretreatment (P<0.05). To further characterize the function of exosomes at the molecular level, protein profiling of exosomes from different cell origins was performed, which identified 129 proteins. Among these, adenylyl cyclase-associated protein 1, a protein implicated in HCC metastasis, was significantly enriched in exosomes from cells with high motile ability (P<0.05). The results of the present study validated the regulatory effect of exosomes on the motile ability of HCC cells. Furthermore, systematic analysis of the protein profiles of exosomes from different origins identified potential factors correlated with HCC metastasis, which may provide a basis for future functional analysis of exosomes regarding their involvement in cancer metastasis and recurrence.

SUBMITTER: Wang S

PROVIDER: S-EPMC5727617 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

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Role of exosomes in hepatocellular carcinoma cell mobility alteration.

Wang Sen S Chen Geng G Lin Xiao X Xing Xiaohua X Cai Zhixiong Z Liu Xiaolong X Liu Jingfeng J

Oncology letters 20171023 6

Exosomes have gained increased research focus due to their key roles as messengers. The components of exosomes include proteins and RNAs that may be horizontally transferred between adjacent or distant cells. Hepatocellular carcinoma (HCC) is among the most malignant types of cancer worldwide, with exosomes implicated to play a crucial role in its regulation; however, the possible function of exosomes in modulating the motile ability of tumor cells and key molecules in HCC remain largely unknown ...[more]

PMID: 29250190

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Project description:We investigated the role of mitochondrial DNA (mtDNA) copy number alteration in human renal cell carcinoma (RCC). The mtDNA copy numbers of paired cancer and non-cancer parts from five resected RCC kidneys after radical nephrectomy were determined by quantitative polymerase chain reaction (Q-PCR). An RCC cell line, 786-O, was infected by lentiviral particles to knock down mitochondrial transcriptional factor A (TFAM). Null target (NT) and TFAM-knockdown (TFAM-KD) represented the control and knockdown 786-O clones, respectively. Protein or mRNA expression levels of TFAM; mtDNA-encoded NADH dehydrogenase subunit 1 (ND1), ND6 and cytochrome c oxidase subunit 2 (COX-2); nuclear DNA (nDNA)-encoded succinate dehydrogenase subunit A (SDHA); v-akt murine thymoma viral oncogene homolog 1 gene (AKT)-encoded AKT and v-myc myelocytomatosis viral oncogene homolog gene (c-MYC)-encoded MYC; glycolytic enzymes including hexokinase II (HK-II), glucose 6-phosphate isomerase (GPI), phosphofructokinase (PFK), and lactate dehydrogenase subunit A (LDHA); and hypoxia-inducible factors the HIF-1? and HIF-2?, pyruvate dehydrogenase kinase 1 (PDK1), and pyruvate dehydrogenase E1 component ? subunit (PDHA1) were analyzed by Western blot or Q-PCR. Bioenergetic parameters of cellular metabolism, basal mitochondrial oxygen consumption rate (mOCRB) and basal extracellular acidification rate (ECARB), were measured by a Seahorse XF(e)-24 analyzer. Cell invasiveness was evaluated by a trans-well migration assay and vimentin expression. Doxorubicin was used as a chemotherapeutic agent. The results showed a decrease of mtDNA copy numbers in resected RCC tissues (p = 0.043). The TFAM-KD clone expressed lower mtDNA copy number (p = 0.034), lower mRNA levels of TFAM (p = 0.008), ND1 (p = 0.007), and ND6 (p = 0.017), and lower protein levels of TFAM and COX-2 than did the NT clone. By contrast, the protein levels of HIF-2?, HK-II, PFK, LDHA, AKT, MYC and vimentin; trans-well migration activity (p = 0.007); and drug resistance to doxorubicin (p = 0.008) of the TFAM-KD clone were significantly higher than those of the NT clone. Bioenergetically, the TFAM-KD clone expressed lower mOCRB (p = 0.009) but higher ECARB (p = 0.037) than did the NT clone. We conclude that a reduction of mtDNA copy number and decrease of respiratory function of mitochondria in RCC might be compensated for by an increase of enzymes and factors that are involved in the upregulation of glycolysis to confer RCC more invasive and a drug-resistant phenotype in vitro.

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Role of exosomes in hepatocellular carcinoma cell mobility alteration.

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Role of exosomes in hepatocellular carcinoma cell mobility alteration.

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