Nc886 promotes renal cancer cell drug-resistance by enhancing EMT through Rock2 phosphorylation-mediated ß-catenin nuclear translocation.

Drug resistance is a significant challenge in the present treatment regimens of renal cell carcinoma (RCC). Our previous study confirmed that nc886 functions as an oncogene in RCC. Nevertheless, the role and underlying mechanism of nc886 in RCC drug resistance are unclear. In the present study, Sunitinib and Everolimus treatment, respectively, downregulated nc886 expression in a dose-dependent manner in all four renal cancer cell lines. Nc886 overexpression in 786-O cells and ACHN cells significantly reduced the sensitivity of cancer cells to both Sunitinib and Everolimus treatment, respectively, by promoting cell viability and inhibiting cell apoptosis, whereas nc886 silencing increased cancer cell sensitivity. In renal cancer cell line with the highest drug-resistance, 786-O cells, Sunitinib, or Everolimus treatment enhanced the cellular EMT and was further enhanced by nc886 overexpression while attenuated by nc886 silencing. In 786-O cells, nc886 overexpression significantly promoted EMT, ROCK2 phosphorylation, and ß-catenin nucleus translocation under Sunitinib or Everolimus treatment. Moreover, ROCK2 silencing significantly reversed the effects of nc886 overexpression on EMT, ROCK2 phosphorylation, and ß-catenin nucleus translocation, as well as drug-resistant renal cancer cell viability and apoptosis. In conclusion, it was demonstrated that nc886 promotes renal cancer cell proliferation, migration, and invasion, as demonstrated previously. nc886 also promotes renal cancer cell drug-resistance to Sunitinib or Everolimus by promoting EMT through Rock2 phosphorylation-mediated nuclear translocation of ß-catenin.

Plasma cell-free DNA and its DNA integrity as biomarker to distinguish prostate cancer from benign prostatic hyperplasia in patients with increased serum prostate-specific antigen.

OBJECTIVES: To investigate whether plasma cell-free DNA (cfDNA) or its integrity could differentiate prostate cancer from benign prostate hyperplasia (BPH) in patients with serum prostate-specific antigen (PSA) ≥ 4 ng/ml. METHODS: Ninety-six patients with prostate cancer and 112 patients with BPH were enrolled. cfDNA levels in plasma before prostate biopsy were quantified by real-time PCR amplification of ALU gene (product size of 115 bp), and quantitative ratio of ALU (247 bp) to ALU (115 bp) reflected the integrity of cfDNA. RESULTS: In patients with serum PSA ≥ 4 ng/ml, there were significant differences in plasma cfDNA or its integrity between the patients with prostate cancer (19.74 ± 4.43, 0.34 ± 0.05) and patients with BPH (7.36 ± 1.58, 0.19 ± 0.03; P < 0.001, P < 0.001). Prostate cancer could be differentiated with a sensitivity of 73.2 % and a specificity of 72.7 % by cfDNA (AUC = 0.864). The integrity of cfDNA had a sensitivity of 81.7 % and a specificity of 78.8 % for the distinguishing prostate cancer from BPH (AUC = 0.910). CONCLUSIONS: cfDNA and its integrity could be applied to differentiate prostate cancer from BPH in patients with serum PSA ≥ 4 ng/ml.

Prediction of clear cell renal cell carcinoma by integrity of cell-free DNA in serum.

OBJECTIVES: To hypothesize that serum cell-free DNA integrity may be clinically useful for prediction of clear cell renal cell carcinoma (cRCC). The integrity of cell-free DNA released from cancer cells was different from that released from apoptotic cells. METHODS: We collected peripheral blood samples from 78 patients before surgery; among these patients, 22 with tumor, both pre- and postoperation, and 42 controls without tumor. After the column extraction of DNA, we performed conventional polymerase chain reaction using different sizes of primers (size of products: 109, 193, 397, and 456 bp) of the housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase for measurement of serum cf-DNA integrity. RESULTS: Age and gender were not associated with cf-DNA integrity in controls (P = .136 and P = .345). Among the cRCC patients, we observed no significant association between cf-DNA integrity and gender, age, tumor grade (P = .510, P = .618 and P = .052), except tumor stage and size (P = .001 and P = .001). All specimens contained 109 bp products. The 193 bp product was detected in 75 of 78 cRCCs and 37 of 42 controls (P = .091), 21 of 22 patients preoperation and 19 of 22 postoperation (P = .294). The 397 bp product was detected in 71 of 78 cRCCs and 0 of 42 of controls (P = .001), 18 of 22 patients preoperation- and 7 of 22 postoperation (P = .003). The 456 bp product was detected in 69 of 78 of cRCCs and 0 of 42 of controls (P = .001), 18 of 22 preoperation and 3 of 22 postoperation (P = .002). CONCLUSIONS: Serum cf-DNA integrity may be a potential tool for the detection of clear cRCC.