Monocarboxylate transporter 1 promotes proliferation and invasion of renal cancer cells by mediating acetate transport.

One hallmark of renal cell carcinoma (RCC) is metabolic reprogramming, which involves elevation of glycolysis and upregulation of lipid metabolism. However, the mechanism of metabolic reprogramming is incompletely understood. Monocarboxylate transporter 1 (MCT1) promotes transport for lactate and pyruvate, which are crucial for cell metabolism. The aim of present study was to investigate the function of MCT1 on RCC development and its mechanism on metabolic reprogramming. The results showed that MCT1 messenger RNA and protein levels significantly increased in cancer tissues of ccRCC compared to normal tissue. MCT1 was further found to mainly located in the cell membrane of RCC. The knockdown of MCT1 by RNAi significantly inhibited proliferation and migration of 786-O and ACHN cells. MCT1 also induced the expressions of proliferation marker Ki-67 and invasion marker SNAI1. Moreover, we also showed that acetate treatment could upregulate the expression of MCT1, but not other MCT isoforms. On the other hand, MCT1 was involved in acetate transport and intracellular histone acetylation. In summary, this study revealed that MCT1 is abnormally high in ccRCC and promotes cancer development. The regulatory effect of MCT1 on cell proliferation and invasion maybe mediated by acetate transport.

miR-520e regulates cell proliferation, apoptosis and migration in breast cancer.

Previous studies have indicated that the deregulation of microRNAs contributes to tumorigenesis. Misregulation of microRNA-520e (miR-520e) has been observed in various types of cancer. However, the expression profile and biological function of miR-520e in breast cancer remains largely unknown. The present study demonstrated that miR-520e expression was significantly increased in breast cancer tissues compared with adjacent non-cancerous breast tissues in 21 patients, as revealed by reverse transcription-quantitative polymerase chain reaction. Furthermore, the proliferation capacity of breast cancer cells was markedly enhanced by the introduction of miR-520e in vitro using a cell counting kit-8 assay. The present study also revealed that the overexpression of miR-520e could suppress breast cancer cell apoptosis, revealed using Annexin V/propidium iodide double staining and flow cytometry analysis. In addition, the ectopic expression of miR-520e promoted the migration of breast cancer cells in vitro, as demonstrated by a Transwell assay. Overall, the findings of the present study highlight an important role for miR-520e in breast cancer development and in the molecular etiology of breast cancer, which indicates the potential application of miR-520e in cancer therapy.

GALNT7, a target of miR-494, participates in the oncogenesis of nasopharyngeal carcinoma.

GalNAc-transferase-7 (GALNT7) is essential for the regulation of cell proliferation and has been implicated in tumorigenesis. However, the role of GALNT7 in the development and progression of nasopharyngeal carcinoma (NPC) remains unclear. Our previous study showed that GALNT7 was a putative target of miR-494, which was confirmed by luciferase reporter assay. In the present study, we demonstrated that in vitro knockdown of GALNT7 significantly inhibited the proliferation, colony formation, migration, and invasion of NPC-derived cells. In vivo tumorigenicity assay showed that miR-494 and GALNT7-small interfering RNA (siRNA) reduced tumor growth in nude mice. Taken together, our results provided new evidence for an oncogenic role of GALNT7 in NPC.

Potential transcriptional regulatory regions exist upstream of the human ezrin gene promoter in esophageal carcinoma cells.

We previously demonstrated that the region -87/+134 of the human ezrin gene (VIL2) exhibited promoter activity in human esophageal carcinoma EC109 cells, and a further upstream region -1324/-890 positively regulated transcription. In this study, to identify the transcriptional regulatory regions upstream of the VIL2 promoter, we cloned VIL2 -1541/-706 segment containing the -1324/-890, and investigated its transcriptional regulatory properties via luciferase assays in transiently transfected cells. In EC109 cells, it was found that VIL2 -1541/-706 possessed promoter and enhancer activities. We also localized transcriptional regulatory regions by fusing 5'- or 3'-deletion segments of VIL2 -1541/-706 to a luciferase reporter. We found that there were three positive and one negative transcriptional regulatory regions within VIL2 -1541/-706 in EC109 cells. When these regions were separately located upstream of the luciferase gene without promoter, or located upstream of the VIL2 promoter or SV40 promoter directing the luciferase gene, only VIL2 -1297/-1186 exhibited considerable promoter and enhancer activities, which were lower than those of -1541/-706. In addition, transient expression of Sp1 increased ezrin expression and the transcriptional activation of VIL2 -1297/-1186. Other three regions, although exhibiting significantly positive or negative transcriptional regulation in deletion experiments, showed a weaker or absent regulation. These data suggested that more than one region upstream of the VIL2 promoter participated in VIL2 transcription, and the VIL2 -1297/-1186, probably as a key transcriptional regulatory region, regulated VIL2 transcription in company with other potential regulatory regions.

Transplantation of microencapsulated umbilical-cord-blood-derived hepatic-like cells for treatment of hepatic failure.

AIM: To investigate intraperitoneal transplantation of microencapsulated hepatic-like cells from human umbilical cord blood for treatment of hepatic failure in rats. METHODS: CD34+ cells in umbilical cord blood cells were isolated by magnetic cell sorting. In the in vitro experiment, sorted CD34+ cells were amplified and induced into hepatic-like cells by culturing with a combination of fibroblast growth factor 4 and hepatocyte growth factor. Cultures without growth factor addition served as controls. mRNA and protein levels for hepatic-like cells were analyzed by reverse transcription-polymerase chain reaction, immunohistochemistry and immunofluorescence. In the in vivo experiment, the hepatic-like cells were encapsulated and transplanted into the abdominal cavity of acute hepatic failure (AHF) rats at 48 h after D-galactosamine induction of acute hepatic failure. Transplantation with PBS and unencapsulated hepatic-like cells served as controls. The mortality rate, hepatic pathological changes and serum biochemical indexes were determined. The morphology and structure of microcapsules in the greater omentum were observed. RESULTS: Human albumin, alpha-fetoprotein and GATA-4 mRNA and albumin protein positive cells were found among cultured cells after 16 d. Albumin level in culture medium was significantly increased after culturing with growth factors in comparison with culturing without growth factor addition (P < 0.01). Compared with the unencapsulated group, the mortality rate of the encapsulated hepatic-like cell-transplanted group was significantly lower (P < 0.05). Serum biochemical parameters, alanine aminotransferase, aspartate aminotransferase and total bilirubin in the encapsulated group were significantly improvement compared with the PBS control group (P < 0.01). Pathological staining further supported these findings. At 1-2 wk post-transplantation, free microcapsules with a round clear structure and a smooth surface were observed in peritoneal lavage fluid, surviving cells inside microcapsules were found by trypan blue staining, but some fibrous tissue around microcapsules was also detected in the greater omentum of encapsulated group by hematoxylin and eosin staining. CONCLUSION: Transplantation of microencapsulated hepatic-like cells derived from umbilical cord blood cells could preliminarily alleviate the symptoms of AHF rats.

[Establishment of animal model for in vitro spermatogenesis by heterotopic grafting of neonatal mouse testis].

OBJECTIVE: To establish an in vitro model for the development of mouse spermatogenic cells into sperm by using the immunodefective mouse as the incubator. METHODS: Tissue grafting was performed using testis from 1-2 days old Kun-ming mice as donor tissue and immunodefective mice as recipients; the expression of TESK1 mRNA in grafts was determined by RT-PCR and the spermatogenesis further observed with histological analysis of grafts. RESULTS: Molecular biological and histological analyses showed that grafts post-grafting not only expressed TESK1 mRNA as in normal mouse testis, but also exhibited similarities in the structure of seminiferous tubules and component of spermatogenic cells, including sperms. CONCLUSION: Spermatogenic cells heterotopically grafted in vitro could continuously grow and complete spermatogenesis and finally develop into sperm.