[Retracted] Downregulation of miR­29b targets DNMT3b to suppress cellular apoptosis and enhance proliferation in pancreatic cancer.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that certain of the Hoechst staining data shown in Fig. 4E were strikingly similar to data appearing in different form in another article by different authors at a different research institute; moreover, an unexpectedly high degree of similarity was noted with the data featured in a couple of different data panels showing the results of apoptosis experiments in Fig. 4D. Owing to the fact that the contentious data in the above article had already been published prior to its submission to Molecular Medicine Reports, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 17: 2113­2120, 2018; DOI: 10.3892/mmr.2017.8145].

Downregulation of miR­29b targets DNMT3b to suppress cellular apoptosis and enhance proliferation in pancreatic cancer.

As one of the most aggressive types of tumor, pancreatic cancer is a principal cause of tumor­associated mortality. Negative associations between microRNA­29 (miR­29) and DNA methyltransferases (DNMT) 3a and 3b have been demonstrated to be associated with the carcinogenesis of a number of types of cancer; however, this has not been completely elucidated in pancreatic cancer. In the present study, pancreatic cancer tissues (n=15) and corresponding paracancerous tissues (n=15) were obtained and the results of reverse transcription­quantitative polymerase chain reaction analysis indicated decreased expression of miR­29b and enhanced mRNA expression of DNMT3b in pancreatic cancer tissues, compared with the corresponding paracancerous tissues. Increased protein expression of DNMT3b was demonstrated by western blotting and immunohistochemistry. In addition, the negative association between miR­29b and DNMT3b was noted in pancreatic cancer tissues, and luciferase reporter assays confirmed that miR­29b was able to directly target DNMT3b in vitro. Notably, miR­29b overexpression was able to decrease cell viability and to promote the apoptosis by targeting DNMT3b, and the knockdown of DNMT3b exhibited consistent results in vitro and in vivo. The results of the present study suggested that miR­29b, as a tumor suppressor, may be a novel target for the development of treatments for pancreatic cancer.

Activated rat hepatic stellate cells influence Th1/Th2 profile in vitro.

AIM: To investigate the effects of activated rat hepatic stellate cells (HSCs) on rat Th1/Th2 profile in vitro. METHODS: Growth and survival of activated HSCs and CD4(+) T lymphocytes cultured alone or together was assessed after 24 or 48 h. CD4(+) T lymphocytes were then cultured with or without activated HSCs for 24 or 48 h and the proportion of Th1 [interferon (IFN)-γ(+)] and Th2 [interleukin (IL)-4(+)] cells was assessed by flow cytometry. Th1 and Th2 cell apoptosis was assessed after 24 h of co-culture using a caspase-3 staining procedure. Differentiation rates of Th1 and Th2 cells from CD4(+) T lymphocytes that were positive for CD25 but did not express IFN-γ or IL-4 were also assessed after 48 h of co-culture with activated HSCs. Galectin-9 expression in HSCs was determined by immunofluorescence and Western blotting. ELISA was performed to assess galectin-9 secretion from activated HSCs. RESULTS: Co-culture of CD4(+) T lymphocytes with activated rat HSCs for 48 h significantly reduced the proportion of Th1 cells compared to culture-alone conditions (-1.73% ± 0.71%; P < 0.05), whereas the proportion of Th2 cells was not altered; the Th1/Th2 ratio was significantly decreased (-0.44 ± 0.13; P < 0.05). In addition, the level of IFN-γ in Th1 cells was decreased (-65.71 ± 9.67; P < 0.01), whereas the level of IL-4 in Th2 cells was increased (82.79 ± 25.12; P < 0.05) by co-culturing, as measured by mean fluorescence intensity by flow cytometry. Apoptosis rates in Th1 (12.27% ± 0.99%; P < 0.01) and Th2 (1.71% ± 0.185%; P < 0.01) cells were increased 24 h after co-culturing with activated HSCs; the Th1 cell apoptosis rate was significantly higher than in Th2 cells (P < 0.01). Galectin-9 protein expression was significantly decreased in HSCs only 24 h after co-culturing (P < 0.05) but not after 48 h. Co-culture for 48 h significantly increased the differentiation of Th1 and Th2 cells; however, the increase in the proportion of Th2 cells was significantly higher than that of Th1 cells (1.85% ± 0.48%; P < 0.05). CONCLUSION: Activated rat HSCs lower the Th1/Th2 profile, inhibiting the Th1 response and enhancing the Th2 response, and this may be a novel pathway for liver fibrogenesis.

Hepatitis B virus particles preferably induce Kupffer cells to produce TGF-ß1 over pro-inflammatory cytokines.

BACKGROUND: Kupffer cells and related cytokines are thought to play a critical role in liver fibrosis; however, the role played by Kupffer cells in hepatitis B virus-related fibrogenesis is unknown. METHODS: Primary rat Kupffer cells were cultured with different titres of hepatitis B virus particles and the concentrations of transforming growth factor (TGF)-ß1, interleukin (IL)-1, IL-6 and tumour necrosis factor (TNF)-α in the culture supernatant were measured every 24h for 7 days. The mRNA and protein levels of these cytokines in Kupffer cells were also analysed using quantitative real-time polymerase chain reaction and western blotting, respectively. RESULTS: Kupffer cells maintained normal morphology and function throughout the 7-day exposure to hepatitis B virus. The concentration of TGF-ß1 secreted by hepatitis B virus-stimulated Kupffer cells (6 log IU/ml hepatitis B virus) increased 5.38- and 7.75-fold by Days 3 and 7, respectively (p<0.01). Western blotting showed that TGF-ß1 expression in Kupffer cells exposed to high titres of hepatitis B virus increased 1.80- and 2.42-fold by Days 3 and 7, respectively (p<0.01). In contrast, Kupffer cell expression and secretion of pro-inflammatory cytokines (IL-6, IL-1 and TNF-α) was unchanged throughout the experiment. CONCLUSION: Hepatitis B virus preferentially stimulates Kupffer cells to produce the pro-fibrogenic/anti-inflammatory cytokine TGF-ß1 rather than the pro-inflammatory cytokines IL-6, IL-1 and TNF-α. This may partly explain why overt liver fibrosis still presents in cases of chronic hepatitis B virus infection with minimal (or no) necro-inflammation.

Rat renal interstitial fibroblasts affect the TH1/TH2 profile in vitro.

BACKGROUND: T helper 1 (Th1)/T helper 2 (Th2) profile is pivotal in the development of fibrosis. Renal interstitial fibroblasts, which play a central role in the development of renal interstitial fibrosis, have an intimate relation with lymphocytes. However, there is little knowledge of the effect of fibroblasts on the profile of CD4 T-lymphocyte subsets. METHODS: After coculture with rat renal interstitial fibroblasts, the proportions of Th1 and Th2 cells in CD4 T lymphocytes and the apoptosis rates of the two subsets were detected by flow cytometry. Galectin-9 expression in rat renal interstitial fibroblasts was detected by immunofluorescence, Western blotting, and enzyme-linked immunosorbent assay. RESULTS: After 48 h of coculture, rat renal interstitial fibroblasts increased the proportion of Th2 cells, lowering the ratio of Th1/Th2. Meanwhile, interferon-gamma production in Th1 cells was inhibited and interleukin-4 production in Th2 was promoted. After coculture with activated rat renal interstitial fibroblasts for 24 h, apoptosis of Th1 was more highly promoted than that of Th2 cells. In addition, rat renal interstitial fibroblasts induced stronger Th2 cell differentiation than that of Th1 cells in vitro. Rat renal interstitial fibroblasts expressed but did not secrete galectin-9 (an apoptosis-inducing factor for Th1 cells) in vitro and the expression level decreased when cocultured with CD4 T lymphocytes. CONCLUSIONS: Rat renal interstitial fibroblasts shift the Th1/Th2 profile in vitro, and this may be another pathway by which renal interstitial fibroblasts promote fibrosis.

[Expression of interleukin-17 in hepatitis B related liver fibrosis].

AIM: To explore the role of interleukin-17 (IL-17) in the pathogenesis and progress of chronic hepatitis B virus (HBV) related liver diseases, especially in hepatitis B related liver fibrosis. METHODS: Whole vein blood was obtained from the patients with chronic hepatitis B (CHB) (n=36), liver cirrhosis (LC) (n=42) including child-pugh A (n=15), child-pugh B (n=12), child-pugh C (n=15), primary hepatitis carcinoma (PHC) (n=34), chronic severe hepatitis (n=30) and normal control (n=20).The level of serum IL-17 and the expression of IL-17 mRNA in peripheral blood mononuclear cell (PBMC) were analysed by ELISA and RT-PCR respectively. Serum IL-17 and liver fibrosis index (C-IV, LN, HA) concentration of LC classified by child-pugh scores were detected by ELISA. RESULTS: The level of serum IL-17 and the expression of IL-17 mRNA in PBMC in four HBV infectious groups were higher than that in control group ( P<0.01). The level of serum IL-17 and the expression of IL-17 mRNA in PBMC in LC group was higher than that in other groups ( P<0.01). The level of serum IL-17 and liver fibrosis index in child-pugh C group in LC patients were higher than that in Child-pugh B group and that in child-pugh B group were higher than that in child-pugh A group (P<0.01). The level of serum IL-17 in LC group positively correlated to child-pugh score and liver fibrosis index (C-IV, LN, HA) (r=0.582, 0.568, 0.682, 0.764, P<0.01). CONCLUSION: IL-17 is obviously increased in HBV related liver diseases, especially in hepatitis B related liver cirrhosis, indicating that IL-17 may play a part in the development of HBV related liver diseases, especially the pathophysiology of hepatitis B related liver fibrosis.