Exosomal miR-4800-3p Aggravates the Progression of Hepatocellular Carcinoma via Regulating the Hippo Signaling Pathway by Targeting STK25.

Background: Emerging evidence has shown that exosome microRNAs (miRNAs) regulate the development of hepatocellular carcinoma (HCC). Here, the influences of miR-4800-3p on the progression of HCC were explored. Materials and Methods: The expression of miR-4800-3p in the exosome derived by transforming growth factor beta 1 (TGF-ß1)-treated HCC cells and the serum exosome isolated from HCC patients were identified by real-time PCR. The effects of TGF-ß1 and the influences of Huh7-secreted exosomes and the effects of miR-4800-3p combined with/without STK25 on cell functions were explored using the EdU assay cloning experiments, wound healing assay, and Transwell assay. The corresponding molecular mechanisms were further detected using Western blot and real-time PCR assays. The combination of miR-4800-3p and STK25 was verified by the dual-luciferase and RNA pulldown assays. The influences of miR-4800-3p on the growth and epithelial-mesenchymal transformation (EMT) of implanted tumors were tested in vivo and further confirmed by Western blot. Results: The miR-4800-3p expression was highly expressed in both exosomes derived by TGF-ß1-treated HCC cells and the serum exosomes of HCC patients. In the cases of treatment with both Huh7-derived exosomes, the level of miR-4800-3p expression was highest, and the treatment of TGF-ß1 could greatly promote the proliferation, stemness, migration, and invasion of HCC cells via upregulating the markers of stemness and EMT, including CD44, CD133, OCT4, N-cadherin, E-cadherin, and ZO-1. Similar results could be obtained when miR-4800-3p was overexpressed in HCC cells. Furthermore, downregulation of STK25 expression, a direct target gene of miR-4800-3p, could greatly rescue the malignant biological behaviors aggravated by overexpression of miR-4800-3p. This was achieved by suppressing the expression of CD44, CD133, OCT4, N-cadherin, and PCNA and activating the Hippo pathway while increasing E-cadherin and ZO-1. Similar results were also obtained in vivo that knockdown of miR-4800-3p expression suppressed tumor growth induced by Huh7-derived exosomes by mediating the EMT markers and the Hippo signaling pathway. Conclusion: Exosomal miR-4800-3p could accelerate HCC development by regulating the Hippo signal by targeting STK25, which could be used as a new therapeutic target for HCC treatment.

The effect of chemokine CC motif ligand 19 on the proliferation and migration of hepatocellular carcinoma.

Multiple studies have shown that CC motif chemokine ligand 19 (CCL19) promotes cell proliferation in several human cancers. The aim of this study was to investigate the expression and function of CCL19 in hepatocellular carcinoma (HCC). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blot, and immunohistochemistry were performed separately to detect the expression of CCL19 in HCC tissues. The expression of CCL19 and its receptor (CCR7) in different HCC cell lines were screened by Western blot. HCC cell lines were screened and processed with recombinant human CCL19 (rhCCL19) or si-CCL19 RNA. Cell proliferation assay and transwell assay were performed to evaluate the proliferation and migration of HCC cells, respectively. Low expression of CCL19 was observed in 83.72 % (72/86) of the HCC versus 16.67 % (4/24) of the adjacent non-tumorous liver tissues, the difference of CCL19 expression between HCC and adjacent non-tumorous liver tissues was statistically significant (P < 0.001). The expression level of CCL19 mRNA and protein in tumor tissues was significantly lower than adjacent non-tumorous liver tissues. The proliferation and migration of HCC cells were obviously inhibited in rhCCL19-treated groups. Our data suggest that CCL19 may play a suppressive role in the regulation of aggressiveness in human HCC.

The influence of g.19124G>a genetic polymorphism in the OPG gene on bone mineral density in Chinese women.

OBJECTIVE: The aim of this study was to detect the influence of g.19124G>A genetic polymorphism in the osteoprotegerin (OPG) gene on bone mineral density (BMD) and osteoporosis in Chinese postmenopausal women. METHODS: A total of 403 primary osteoporosis subjects and 409 healthy controls were enrolled in this study. The BMD value of the femoral neck hip, lumbar spine (L(2-4)), and total hip was analyzed by Norland XR-46 dual energy X-ray absorptiometry. The polymerase chain reaction-restriction fragment length polymorphism method was utilized to investigate the genotype of g.19124G>A genetic polymorphism. RESULTS: We found significant differences of the femoral neck hip, lumbar spine (L(2-4)), and total hip of BMD among different genotypes of g.19124G>A genetic polymorphism, individuals with the genotype GG had significantly higher BMD than those of genotype GA and AA (p<0.05). CONCLUSION: These findings indicate that the g.19124G>A genetic polymorphism in the OPG gene is potentially related to BMD and osteoporosis in Chinese postmenopausal women, and the allele A could be associated with a lower BMD and an increased risk factor for osteoporosis.

Polymorphisms in DNA repair genes and risk of glioma and meningioma.

Polymorphisms in DNA repair genes have been shown to influence DNA repair processes and to modify cancer susceptibility. Here we conducted a case-control study to assess the role of potential SNPs of DNA repair genes on the risk of glioma and meningioma. We included 297 cases and 458 cancer-free controls. Genotyping of XRCC1 Gln399Arg, XRCC1 Arg194Trp, XRCC2 Arg188His, XRCC3 Thr241Met, XRCC4 Ala247Ser, ERCC1 Asn118Asp, ERCC2 Lys751Gln and ERCC5 Asp1558His were performed in a 384-well plate format on the Sequenom MassARRAY platform. XRCC1 Arg194Trp (rs1799782) and ERCC2 Asp312Asn rs1799793 did not follow the HWE in control group, and genotype distributions of XRCC1 Gln399Arg rs25487, XRCC2 Arg188His rs3218536 and ERCC2 Asp312Asn rs1799793 were significantly different between cases and controls (P<0.05). We found XRCC1 399G/G, XRCC1 194 T/T and XRCC3 241T/T were associated with a higher risk when compared with the wild-type genotype. For ERCC5 Asp1558His, we found G/G genotype was associated with elevated susceptibility. In conclusion, our study has shown that XRCC1 Gln399Arg, XRCC1 Arg194Trp, XRCC3 Thr241Met and ERCC5 Asp1558His are associated with risk of gliomas and meningiomas. This finding could be useful in identifying the susceptibility genes for these cancers.