Prognostic effect of DNA methylation of BTG2 gene in Chinese hepatocellular carcinoma.

Background: This study aims to develop a prognostic model for overall survival based on potential methylation sites within B-cell translocation gene 2 (BTG2) in Chinese patients with hepatocellular carcinoma (HCC). Methods: This is a retrospective study. The beta values of nine CpG sites and RSEM normalized count values of BTG2 gene were extracted from the Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) (TCGA-LIHC) dataset, with the beta value representing the methylation level by indicating the ratio of the intensity of the methylated bead type to the combined locus intensity. Pyrosequencing was performed to determine the range of methylation values surrounding cg01798157 site in BTG2 gene. A weighted linear model was developed to predict the overall survival (OS). Results: The beta value of cg01798157 was significantly negatively associated with the mRNA expression of BTG2 in the TCGA-LIHC dataset (Spearman's rho = -0.5306, P = 2.27 × 10-27). The methylation level of cg01798157 was significantly associated with OS in the cohort of 51 Chinese HCC patients (Hazard ratio = 0.597, 95% CI: 0.434-0.820, P = 0.001). Multivariate Cox regression analysis identified methylation level of cg01798157, cirrhosis, and microvascular invasion as independent prognostic factors. The prognostic efficiency of death risk score was superior to that of cirrhosis or microvascular invasion alone. Conclusions: The methylation level of cg01798157 in BTG2 may be an epigenetic biomarker in Chinese patients with resectable HCC.

Pathological complete remission in ALK-positive lung cancer patient after multiple lines of conversion therapy.

Introduction: Traditional therapeutic approaches for the treatment of advanced non-small-cell lung cancer (NSCLC) are based on chemotherapy. However, the discovery and understanding of oncogenic driver alterations has led to the development of targeted therapies that have substantially improved patient outcomes. Still, to date, there have been no reports of patients with advanced anaplastic lymphoma kinase (ALK)-positive lung cancer achieving clinical complete response (cCR) in the systemic lesion and pathological complete remission (pCR) in primary lung lesion after multiple lines of conversion therapy. Methods: In this case, a 55-year-old man was diagnosed with ALK-positive, stage IV lung adenocarcinoma using immunohistochemistry and next generation sequencing (NGS) tests. Results: Crizotinib and two other ATP-competitive ALK inhibitors, ceritinib and alectinib, were used respectively as first-line, second-line, and third-line therapy. The patient received treatment with crizotinib and achieved partial response (PR), but 5 months later the efficacy was evaluated as progressive disease (PD). Ceritinib was used as the second-line treatment, but the disease progressed 6 months later. Alectinib was used as the third-line treatment, but the efficacy was evaluated as PD. From April 2019 to November 2019, the patient received 4 cycles of induction chemotherapy with pemetrexed/carboplatin/bevacizumab and then switched to pemetrexed/bevacizumab as the fourth-line treatment, and received the fifth line treatment, cetuximab/paclitaxel liposome/nedaplatin, for 1 cycle, but the disease still progressed. Then the patient received the sixth line of treatment, camrelizumab/lorlatinib, for 9 antitumor cycles, resulting in PR. The patient underwent surgery followed by maintenance treatment with lorlatinib and achieved cCR. To our knowledge, this is the first documented case of cCR in a patient with ALK-positive advanced lung adenocarcinoma treated with multiple lines of therapy followed by surgical treatment. Discussion: This case reveals the possible survival benefit of immunotherapy after multiple line treatment in ALK-positive advanced lung adenocarcinoma, indicating that it is possible find new therapeutic targets based on NGS molecular detection and provide precise therapeutic strategies for clinical practice when drug resistance or progression occurs in cancer therapy.

Ribonucleic acid-binding protein CPSF6 promotes glycolysis and suppresses apoptosis in hepatocellular carcinoma cells by inhibiting the BTG2 expression.

Hepatocellular carcinoma (HCC) is currently the sixth most common malignancy and the second major cause of tumor-related deaths in the world. This study aimed to investigate the role of cleavage and polyadenylation factor-6 (CPSF6) and B-cell translocation gene 2 (BTG2) in regulating the glycolysis and apoptosis in HCC cells. The RNA and protein expression of CPSF6 and BTG2 in normal hepatocyte and HCC were, respectively, detected by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis and Western blot analysis. The viability and apoptosis of transfected Huh-7 cells were, respectively, analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay. The expression of apoptosis-related proteins and HK-2 in transfected Huh-7 cells was also detected by Western blot analysis. The levels of glucose and lactate in the culture supernatant of transfected Huh-7 cells were, respectively, detected with the glucose assay kit and lactate assay kit. The interaction of CPSF6 and BTG2 was confirmed by RNA binding protein immunoprecipitation (RIP) assay. As a result, CPSF6 expression was increased while BTG2 expression was decreased in Huh-7 cells. Interference with CPSF6 suppressed the viability and glycolysis, and promoted the apoptosis of Huh-7 cells. Furthermore, CPSF6 interacted with BTG2 and interference with CPSF6 upregulated the BTG2 expression and inhibited the protein kinase B (AKT)/extracellular signal-regulated kinase (ERK)/nuclear factor (NF)-κB pathway. Interference with BTG2 could partially reverse the above cell changes caused by interference with CPSF6. In conclusion, CPSF6 inhibited the BTG2 expression to promote glycolysis and suppress apoptosis in HCC cells by activating AKT/ERK/NF-κB pathway.

High expression of c-Met and EGFR is associated with poor survival of patients with glottic laryngeal squamous cell carcinoma.

The present study was undertaken to explore the association between the expression of hepatocyte growth factor receptor (c-Met) and epidermal growth factor receptor (EGFR) with clinicopathological factors and survival status, to obtain prognostic biomarkers in patients with glottis laryngeal squamous cell carcinoma (GLSCC). The expression status of c-Met and EGFR protein was analyzed in 71 archival laryngeal cancer samples by immunohistochemistry. Statistical methods, including univariate and multivariate Cox regression analysis, were used to determine risk factors of progression. In addition, survival analysis was performed by the Kaplan-Meier method. The present study detected positive expression of c-Met and EGFR in 69.0 and 91.5% of GLSCC samples, respectively. The median disease-free survival (DFS) and overall survival (OS) times of all patients were 42.4 and 81.8 months, respectively, and the 2-year DFS and OS rates were 60.1 and 84.91%, respectively. Univariate Cox regression analysis revealed that patients with high expression of EGFR or c-Met had a predisposition for tumor recurrence. The expression of c-Met expression was significantly associated with that of EGFR (P=0.001). High expression of c-Met or EGFR was associated with shorter DFS and OS times. Findings of the multivariate Cox regression analysis indicated that c-Met-expression may be used as an independent predictor of DFS and OS (P=0.002 and P=0.008, respectively). However, EGFR expression was not an independent predictor for DFS and OS (P=0.352 and P=0.24, respectively). The high expression of c-Met and EGFR was associated with poor survival and are important predictors for prognosis of patients with GLSCC.

Target migration from re-inflation of adjacent atelectasis during lung stereotactic body radiotherapy.

Stereotactic body radiotherapy (SBRT) is a widely accepted option for the treatment of medically inoperable early-stage non-small cell lung cancer (NSCLC). Herein, we highlight the importance of interfraction image guidance during SBRT. We describe a case of early-stage NSCLC associated with segmental atelectasis that translocated 15 mm anteroinferiorly due to re-expansion of the adjacent segmental atelectasis following the first fraction. The case exemplifies the importance of cross-sectional image-guided radiotherapy that shows the intended target, as opposed to aligning based on rigid anatomy alone, especially in cases associated with potentially "volatile" anatomic areas.

Expression of B-cell translocation gene 2 is associated with favorable prognosis in hepatocellular carcinoma patients and sensitizes irradiation-induced hepatocellular carcinoma cell apoptosis in vitro and in nude mice.

B-cell translocation gene 2 (BTG2) proteins have been reported to be putative tumor suppressors in various cancer types. The present study first assessed BTG2 expression in 44 human liver cancer tissue specimens, then investigated BTG2 expression in the regulation of hepatocellular carcinoma (HCC) cell apoptosis with or without radiotherapy in vitro and in vivo. The results revealed that BTG2 protein expression was significantly reduced in HCC tissues, and associated with better survival for HCC patients (P=0.05). BTG2 overexpression also sensitized Huh7 cells to radiation-induced apoptosis in vitro and in a nude mouse model, although restoration of BTG2 expression per se did not affect the viability and apoptosis of HCC cells. Future studies would confirm the role of BTG2 in hepatoma, and further develop BTG2 as a therapeutic strategy for controlling HCC.

One-Carbon Metabolic Factors and Risk of Renal Cell Cancer: A Meta-Analysis.

BACKGROUND: Nutrients related to one-carbon metabolism were previously shown to be significantly associated with the risk of cancer. The aim of this meta-analysis was to evaluate potential relationships between one-carbon metabolic factors and renal cell cancer (RCC) risk. METHODS: PubMed, EMBASE, and Cochrane Library databases were searched through March 2015 for observational studies of quantitative RCC risk estimates in relation to one-carbon metabolic factors. The relative risks (RRs) with 95% confidence intervals (CIs) measured the relationship between one-carbon metabolic factors and RCC risk using a random-effects model. RESULTS: Of the 463 citations and abstracts identified by database search, seven cohorts from five observational studies reported data on 133,995 individuals, and included 2,441 RCC cases. Comparing the highest with the lowest category, the pooled RRs of RCC were 0.72 (95%CI: 0.52-1.00; P = 0.048) for vitamin B12. In addition, an increase in folic acid supplementation of 100 µg/day was associated with a 3% lower risk of RCC (RR, 0.97; 95%CI: 0.93-1.00; P = 0.048). Similarly, an increase of 5 nmol/L of vitamin B2 was associated with a reduced risk of RCC 0.94 (95%CI: 0.89-1.00; P = 0.045). Sensitivity analyses suggested that a higher serum vitamin B6 might contribute to a reduced risk of RCC (RR, 0.83; 95%CI: 0.77-0.89; P < 0.001). CONCLUSIONS: Higher levels of serum vitamin B2, B6, B12, and folic acid supplementation lowered the risk of RCC among the study participants.

MicroRNAs involved with hepatocellular carcinoma (Review).

Hepatocellular carcinoma (HCC) is one of the most common malignancies, which accounts for 90% of primary liver cancer. HCC usually presents with poor outcomes due to the high rates of tumor recurrence and widespread metastasis. However, the underlying mechanism of HCC initiation and progression, which significantly hindered the development of valid approaches for early detection and treatment remain to be elucidated. As a group of small non-coding RNAs, microRNAs (miRNAs) have been demonstrated to be involved in many types of diseases especially human malignancies. Numerous miRNAs are deregulated in HCC, which may shed some light on current investigations. Since miRNAs are stable and detected easily, their ectopic expression has been reported in HCC tissues, serum/plasma and cell lines. As previously described, miRNAs serve as tumor suppressors or oncogenes, indicating that miRNAs may be useful as diagnostic, therapeutic and prognostic markers of HCC. In the present review, we assessed the latest data regarding dysregulated miRNAs in HCC and reviewed the reported functions of these miRNAs as they apply to the diagnosis and prognosis of HCC.

MicroRNA­21 regulates the expression of BTG2 in HepG2 liver cancer cells.

B­cell translocation gene 2 (BTG2) is a tumor suppressor gene, which belongs to the anti­proliferation gene family. Our previous study demonstrated that microRNA (miR)­21 and the expression of BTG2 were negatively correlated during hepatocarcinogenesis. The aim of the present study was to investigate the effects of miR­21 on the growth and progression of liver cancer cells, and to determine the underlying mechanism. A luciferase reporter assay was used to demonstrate that the BTG2 gene was a direct target of miR­21. In addition, the effects of miR­21 on cell growth and gene expression in HepG2 human hepatocellular carcinoma (HCC) cells were analyzed using reverse transcription­quantitative polymerase chain reaction, western blotting, an MTT assay, flow cytometry, a Transwell invasion assay and a wound healing assay. The expression levels of miR­21 in the HepG2 cells were significantly higher, compared with those in L02 normal liver cells. The expression levels of BTG2 in liver cancer cell lines (HepG2 and Huh7) were significantly lower, compared with that in the L02 cells. These results suggested that BTG2 was the direct target gene of miR­21. The protein expression levels of BTG2 were inhibited by high expression levels of miR­21, and increased by inhibition of the expression of miR­21 in the HepG2 cells. Inhibition of miR­21 reduced cell proliferation and invasion, and increased the rate of apoptosis in the HepG2 cells. These results indicated that miR­21 regulates cell proliferation, invasion, migration and apoptosis in HepG2 cells, which may be associated with its effects on the expression of BTG2. The results of the present study may provide a basis for targeting the miR­21/BTG2 interaction for the treatment of HCC.

BTG2: a rising star of tumor suppressors (review).

B-cell translocation gene 2 (BTG2), the first gene identified in the BTG/TOB gene family, is involved in many biological activities in cancer cells acting as a tumor suppressor. The BTG2 expression is downregulated in many human cancers. It is an instantaneous early response gene and plays important roles in cell differentiation, proliferation, DNA damage repair, and apoptosis in cancer cells. Moreover, BTG2 is regulated by many factors involving different signal pathways. However, the regulatory mechanism of BTG2 is largely unknown. Recently, the relationship between microRNAs and BTG2 has attracted much attention. MicroRNA-21 (miR-21) has been found to regulate BTG2 gene during carcinogenesis. In this review, we summarize the latest findings in the investigations of biological functions of BTG2 and regulation of its expression, with an emphasis on miR-21 in regulation of BTG2 gene in various cancers. B-cell translocation gene 2 (BTG2), also known as PC3 or TIS21, belongs to the antiproliferative (APRO) gene family. Several studies have demonstrated that BTG2 is involved in a large number of physiological and pathological processes, such as cell differentiation, proliferation, apoptosis, and other cellular functions, acting as a tumor suppressor. In this review, we summarize the latest findings in BTG2 studies, highlighting the mechanisms for the regulatory effects of microRNAs (miRNAs) on BTG2 gene expression in the most common human cancers.

miR-224 promotion of cell migration and invasion by targeting Homeobox D 10 gene in human hepatocellular carcinoma.

BACKGROUND AND AIM: MicroRNAs (miRNAs) are small noncoding RNA molecules that control target gene expression and are implicated in the regulation of diverse cellular pathways. In our previous research, we have demonstrated that miR-224 was overexpressed in liver cancer cells and tissues, which was an important factor in the regulation of cell migration and invasion. This study aimed to further explore the regulatory mechanism of miR-224 in the migration and invasion in liver cancer cells. METHODS: A luciferase reporter assay was used to confirm that the HOXD10 gene was a direct target of miR-224. Quantitative reverse transcriptase-polymerase chain reaction, Western blotting, Transwell migration, and Matrigel invasion assays were performed to clarify the molecular mechanism of miR-224 in the regulation of cell migration and invasion in human hepatocellular carcinoma (HCC). RESULTS: (i) The expression of miR-224 was strongly upregulated in MHHC97H and MHCC97L cells, and its expression level was significantly associated with cell invasive potential. (ii) The HOXD10 gene was confirmed to be a direct target of miR-224. Compared with normal liver tissues and cells, HOXD10 had lower expression in HCC tissues and cells and inversely regulated HCC cell invasion. (iii) miR-224 promoted expression of the tumor invasion-associated proteins p-PAK4 and MMP-9 by directly targeting HOXD10. CONCLUSION: Our findings suggest a previously undescribed regulatory pathway in which the miR-224/HOXD10/p-PAK4/MMP-9 signaling pathway contributes to the regulation of cell migration and invasion and provides a new biotarget for HCC treatment.