CDK4/6 inhibition sensitizes MEK inhibition by inhibiting cell cycle and proliferation in pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is not sensitive to most chemotherapy drugs, leading to poor chemotherapy efficacy. Recently, Trametinib and Palbociclib have promising prospects in the treatment of pancreatic cancer. This article aims to explore the effects of Trametinib on pancreatic cancer and address the underlying mechanism of resistance as well as its reversal strategies. The GDSC (Genomics of Drug Sensitivity in Cancer) and CTD2 (Cancer Target Discovery and Development) were utilized to screen the potential drug candidate in PDAC cell lines. The dose-increase method combined with the high-dose shock method was applied to induce the Trametinib-resistant PANC-1 and MIA PaCa-2 cell lines. The CCK8 proliferation assay, colony formation assay, flow cytometry, and western blot were conducted to verify the inhibitory effect of Trametinib and Palbociclib. RNA-seq was performed in resistant PDAC cell lines to find the differential expression genes related to drug resistance and predict pathways leading to the reversal of Trametinib resistance. The GDSC and CTD2 database screening revealed that Trametinib demonstrates a significant inhibitory effect on PDAC. We found that Trametinib has a lower IC50 than Gemcitabine in PDAC cell lines. Both Trametinib and Gemcitabine can decrease the proliferation capacity of pancreatic cells, induce cell cycle arrest, and increase apoptosis. Simultaneously, the phosphorylation of the AKT and ERK pathways were inhibited by the treatment of Trametinib. In addition, the RNA-seq of Trametinib-induced resistance PDAC cell lines reveals that the cyclin-dependent kinase (CDK)-RB-E2F regulatory axis and G2/M DNA damage checkpoint might lead the drug resistance. Besides, the combination of Trametinib with Palbociclib could inhibit the proliferation and cell cycle of both resistant cells lines and also restore the sensitivity of drug-resistant cells to Trametinib. Last but not least, the interferon-α and interferon-γ expression were upregulated in resistance cell lines, which might lead to the reversal of drug resistance. The study shows Trametinib has a critical inhibitory effect on PDAC. Besides, the combination of Trametinib with Palbociclib can inhibit the proliferation of PDAC-resistant cells.

Real-time fluorescence-guided adhesiolysis with indocyanine green in intra-abdominal surgery (with video).

Intra-abdominal adhesions have consistently posed a challenge for surgeons during procedures. This study aims to investigate the feasibility of utilizing indocyanine green (ICG) in conjunction with near-infrared imaging for the detection of intra-abdominal adhesions. In vitro, we analyzed factors affecting ICG fluorescence. We divided SD rats into groups to study ICG excretion in different digestive tract regions. Additionally, we reviewed surgical videos from previous cholecystectomy cases, categorizing them by ICG injection timing and assessing fluorescence imaging in various digestive tract regions. Finally, we preoperatively injected ICG into two cholecystectomized patients with abdominal adhesions, guiding intraoperative adhesiolysis with near-infrared fluorescence imaging. In vitro, we observed a significant influence of protein and ICG concentrations on ICG fluorescence intensity. Our rat experiments unveiled a strong and highly significant correlation (Kendall's tau-b = 1, P < 0.001) between the timing of ICG injection and the farthest point of intestinal fluorescence. A retrospective case analysis further validated this finding (Kendall's tau-b = 0.967, P < 0.001). Under the guidance of fluorescence navigation, two cholecystectomized patients with intra-abdominal adhesions successfully underwent adhesiolysis, and no postoperative complications occurred. The intraoperative combination of ICG with near-infrared fluorescence imaging effectively enhances the visibility of the liver, bile ducts, and various segments of the gastrointestinal tract while providing real-time navigation. This real-time fluorescence guidance has the potential to aid surgeons in the dissection of intra-abdominal adhesions.

Real-time fluorescent cholangiography with indocyanine green in laparoscopic cholecystectomy: a randomized controlled trial to establish the optimal indocyanine green dose within 30 min preoperatively.

PURPOSE: To establish the optimal dose of indocyanine green (ICG) to administer intravenously 30 min before laparoscopic cholecystectomy (LC). METHODS: In this randomized controlled trial (RCT), patients undergoing LC for cholecystitis, cholelithiasis, and/or cholecystic polyps were randomized into four groups given four different ICG doses (0.025, 0.1, 0.25, 2.5 mg). Using OptoMedic endoscopy combined with a near-infrared fluorescent imaging system, we evaluated the fluorescence intensity (FI) of the common bile duct and liver at three timepoints: before surgical dissection of the cystohepatic triangle, before clipping of the cystic duct, and before closure. The bile duct-to-liver ratio (BLR) of the FI was analyzed to assess the cholangiography effect. RESULTS: Sixty-four patients were allocated to one of four groups, with 40 patients included in the final analysis. Generally, with increasing ICG doses, the levels of FI in the bile duct and liver increased gradually at each of the three timepoints. Before surgical dissection of the cystohepatic triangle, 0.1-mg ICG showed the highest BLR (F = 3.47, p = 0.0259). Before clipping the cystic duct and before closure, the 0.025- and 0.1-mg groups showed a higher BLR than the 0.25- and 2.5-mg groups (p < 0.05). When setting the ideal cholangiography at a BLR ≥ 1, ≥ 3, or ≥ 5, the 0.1-mg group showed the highest qualified case number at the three timepoints. CONCLUSIONS: The intravenous administration of 0.1-mg ICG, 30 min before LC, is significantly better for fluorescent cholangiography of the extrahepatic biliary structures before dissection and clipping of the cystohepatic triangle. TRIAL REGISTRATION: This study was registered in the Chinese Clinical Trial Registry (ChiCTR) (ChiCTR2200057933).

Role of C14orf166 in viral infection and RNA metabolism and its relationship with cancer (Review).

Chromosome 14 open reading frame 166 (C14orf166) encodes a 28­kDa nuclear and cytoplasmic protein that is involved in viral infection, RNA metabolism, and centrosome structure. It binds to the polymerase acidic protein subunit of the influenza A virus, which is associated with several transcription factors, RNA polymerase II, to activate transcription initiation and promote virus infection. It also interacts with a mature hepatitis C virus core protein to regulate the infection process. In physiological conditions, C14orf166 associates with the proteins, DDX1, HSPC117 and FAM98B, and regulates RNA metabolism and fate. In addition, C14orf166 is overexpressed in a variety of cancer types. Upregulation of C14orf166 may contribute toward cancer malignancy through its impact on glycogen synthase kinase 3ß­mediated signaling, the downregulation of retinoblastoma protein, or the upregulation of IL­6. Therefore, C14orf166 could be used as a biomarker for the diagnosis and prognosis of various cancer types. This review summarized the existent literature about C14orf166, focusing on its functions in physiological and pathological situations.

Extrahepatic biliary tract visualization using near-infrared fluorescence imaging with indocyanine green: optimization of dose and dosing time.

BACKGROUND: The dose and dosing time of indocyanine green (ICG) vary among fluorescence cholangiography (FC) studies. The purpose of this prospective, randomized, exploratory clinical trial was to optimize the dose and dosing time of ICG. METHODS: PubMed was searched to determine the optimal dose. To optimize the dosing time of ICG, a clinical trial was designed with two parts. The first part included patients with T tubes for more than 1 month. After the patient was injected with ICG, bile was collected at 10 time points to explore the change and trends of bile fluorescence intensity (FI). In addition, the results of the first experiment were used to setup a randomized controlled trial (RCT) that aimed to find the optimal dosing timing for ICG injections for laparoscopic cholecystectomy (LC). During surgery, imaging data were collected for analysis. RESULTS: After performing a systematic review, the ICG injection dose for each patient in the clinical trial was 10 mg. Five patients were included in the first part of the study. Bile collected 8 h after ICG injection had a higher FI than bile collected at other time points (p < 0.05), and the FI of bile collected 20 h after ICG injection was nearly zero. In the second part of the experiment, 4 groups of patients (6 patients per group) were injected with 10 mg ICG at 8, 10, 12 and 14 h prior to surgery. The distribution of bile duct FI (p = 0.001), liver FI (p < 0.001), and common bile duct (CBD)-to-liver contrast (p = 0.001) were not the same in each group. Further analysis with the Bonferroni method revealed the following: (1) the FI of the CBD in the 8 h group was significantly different from that in the 14 h group (adjusted p < 0.001); (2) the liver FI of the 8 h group was higher than that of the 10 h group (adjusted p = 0.042) and the 14 h group (adjusted p < 0.001); and (3) the CBD-to-liver contrast of the 8 h group was lower than that of the 10 h group (adjusted p = 0.013) and the 14 h group (adjusted p = 0.001). CONCLUSION: ICG FC enables the real-time identification of extrahepatic bile ducts. The optimal effect of FC can be achieved by performing 10 mg ICG injections 10 to 12 h prior to surgery.

Bone marrow mesenchymal stromal cells attenuate liver allograft rejection may via upregulation PD-L1 expression through downregulation of miR-17-5p.

Studies have reported that bone marrow mesenchymal stem cells (BMSCs) play an important role in immune regulation after organ transplantation. Some of the BMSC-mediated regulatory mechanisms are related to PD-L1 expression. However, the regulatory mechanism of PD-L1 expression is not fully understood. In this experiment, we confirmed the regulatory role of BMSCs in the rejection of liver transplantation and explored the possible mechanism by which PD-L1 expression is regulated in BMSCs. An orthotopic liver transplantation model in rats was established based on the "double-cuff technique". Third-generation BMSCs were injected into the rejection model rats via portal vein. At the same time, sera were obtained from rats in the allograft, isograft and sham-operated groups. The sera from these groups were separately added to BMSCs complete medium to partially mimic the in vivo environment in which BMSCs are exposed. After predicting and analysing microRNAs that regulate PD-L1 expression, we examined the microRNA and PD-L1 expression in BMSCs of the three groups cultured in the conditioned media. Moreover, a luciferase reporter assay was used to confirm the interaction between PD-L1 and microRNA. The study found that the liver function and liver graft histopathology of the BMSC-treated group were better than those of the allograft group. The Kaplan-Meier survival curve analysis showed that the median survival time (MST) of the BMSC-treated group was longer than that of the allograft group. Bioinformatics prediction and analysis showed that miR-17-5p is highly likely to regulate PD-L1. Compared with the other two groups of cells, BMSCs cultured with serum from allograft models showed higher PD-L1 expression, but lower miR-17-5p expression. Pearson correlation analysis showed that miR-17-5p and PD-L1 expression was negatively correlated, and further luciferase reporter assays confirmed that miR-17-5p interacted directly with the 3´-untranslated region (UTR) of PD-L1 mRNA. The results of this study demonstrate that BMSCs can attenuate liver allograft rejection and provide us with the idea that BMSCs may further upregulate PD-L1 expression by downregulating miR-17-5p expression, thereby attenuating liver allograft rejection.

Prognostic value of miR-17-5p in cancers: a meta-analysis.

BACKGROUND: Studies have shown that miR-17-5p plays an important role in the development of cancer. The aim of this meta-analysis was to quantitatively analyze the association of miR-17-5p with prognosis in various cancers. MATERIALS AND METHODS: We searched the PubMed, EMBASE, Web of Science, and Cochrane library databases for relevant studies through August 2017. The prognostic data and clinico-pathological features of overall survival (OS) and disease-free survival (DFS) were extracted to investigate the association between miR-17-5p expression and tumor prognosis. In addition, odds ratios (ORs) were used to assess the correlations between miR-17-5p expression and clinicopathological characteristics. RESULTS: A total of ten studies were incorporated into this systematic review, and we found that high miR-17-5p expression can predict poor OS for malignancies (combined hazard ratio [HR]=1.87; 95% confidence interval [CI], 1.37-2.55; P=0.000) as well as poor DFS (combined HR=1.60; 95% CI, 1.05-2.44; P=0.027). Further subgroup analyses suggested that high miR-17-5p expression was related to poor OS in Asian patients (combined HR=1.92; 95% CI, 1.37-2.71; P=0.000) and the serum/plasma sample source subgroup (combined HR=2.13; 95% CI, 1.36-3.31; P=0.001). The combined OR indicated that the expression of miR-17-5p was associated with lymph node invasion (OR=1.28; 95% CI, 1.05-1.56; P=0.016) and venous invasion (OR=1.92; 95% CI, 1.40-2.63; P=0.000). CONCLUSION: Elevated expression of miR-17-5p suggested a poor prognosis in cancer patients and may serve as a new tumor marker to monitor cancer development and progression.