Genomic based analyses reveal unique mutational profiling and identify prognostic biomarker for overall survival in Chinese small-cell lung cancer.

OBJECTIVE: As an aggressive subtype of lung cancer, small-cell lung cancer (SCLC) presents a poor prognosis. Although molecular and clinical characteristics have been established for SCLC, limited investigation has been performed for predicting survival of SCLC patients. METHODS: Genomic alterations were profiled in Chinese SCLC patients (N = 37) using targeted sequencing. Clonal mutation burden (CMB) integrated the number of mutations with the clonal structure of the tumor. Specific pathways involving DNA damage repair (DDR) and cell cycle as well as CMB were studied as potential biomarkers for prognosis of SCLC. RESULTS: TP53 and RB1 gene mutations were the most common alterations (91.9% and 83.8%, respectively), followed by LRP1B, FAM135B, SPTA1, KMT2D, FAT1, and NOTCH3. Survival analysis revealed that mutation status of the DDR pathway was associated with worse OS in our cohort. Importantly, patients with higher CMB exhibited worse OS in our cohort and this observation was successfully validated in the cBioportal cohort. Moreover, multivariate analysis demonstrated CMB as a promising independent prognostic factor for OS in Chinese SCLC patients. Interestingly, patients with loss of function of RB1, validated by immunohistochemistry staining, appeared to have worse OS. CONCLUSIONS: The mutational profiling of Chinese SCLC patients signified an ethnicity dependent component. CMB was firstly found to be associated with OS of Chinese SCLC patients and could be regarded as a prognostic marker for SCLC.

Analysis of the function and mechanism of DIRAS1 in osteosarcoma.

BACKGROUND: Osteosarcoma is a prevalent malignant bone tumor with a tendency to metastasize to the lungs. In this study, we intend to detect the function and mechanism of DIRAS family GTPase 1 (DIRAS1) in osteosarcoma cells. METHODS: Expression level of DIRAS1 in osteosarcoma cells was analyzed by western blot. Cell location of DIRAS1 in osteosarcoma cells was detected by immunofluorescence. Small interfering RNAs (siRNA)-DIRAS1 and pcDNA3.1-DIRAS1 were employed to regulate DIRAS1 expression. The malignant behaviors of osteosarcoma cells were examined by cell counting kit-8, colony formation, transwell, and wound healing assays. The expression of related proteins was measured by western blot. ELISA and dot blot assays were used to detect the methylation level of m6A. Rescue assays were performed to detect the function of METTL3/METTL14 and DIRASI on osteosarcoma cells. RESULTS: DIRAS1 was located in the nucleus of osteosarcoma cells. Silencing of DIRAS1 in MG63 cells strengthened the proliferation, invasion and migration abilities, as well as blocked the apoptosis ability. Also, p-ERK expression was regulated by DIRAS1 expression, while p-AKT was not affected. Furthermore, DIRAS1 expression was suppressed by METTL3 or/and METTL14 treatment. Moreover, the inhibitory effect of DIRAS1 overexpression on HOS cells malignant behaviors can be reversed by METTL3 and METTL14 joint treatment. The reduced expression of p-ERK induced by DIRAS1 overexpression can be inversed by METTL3 and METTL14 co-treatment. CONCLUSIONS: Taken together, our findings illustrated that DIRAS1 regulated by METTL3 and METTL14 can obviously modulate the malignant behaviors of osteosarcoma cells by inactivating ERK pathway.

The New Concept of Physiological "Riolan's Arch" and the Reconstruction Mechanism of Pathological Riolan's Arch After High Ligation of the Inferior Mesenteric Artery by CT Angiography-Based Small Vessel Imaging.

BACKGROUND: Due to the complexity of anatomical relationship between superior mesenteric artery (SMA) and left colic artery (LCA), there is no unified anatomical concept of "Riolan's arch." There is no consensus as to tie off the inferior mesenteric artery (IMA) at its origin or just below the origin of the left colic artery during radical surgery of sigmoid colon and rectal cancers. The aim of the study is to investigate the anatomy of shortcut anastomotic branches (adjacent branches) of SMA at splenic flexure and to explore how the shortcut pathway (Riolan's arch) was formed, as the compensation of anastomotic branches between MCA and LCA under pathological conditions and the reconstruction and the mechanism of pathological Riolan's arch after high ligation of the inferior mesenteric artery. METHODS: Between January 2018 and May 2020, patients with colorectal cancer who underwent CTA before surgery were enrolled in the study. The anatomy of shortcut anastomotic branch of SMA and LCA was investigated by volume rendering technique (VR) and maximum-intensity projection (MIP). GE's small vessel extraction technology (selected VR) was used to directly display these shortcut anastomotic branches on a map and to establish their three-dimensional anatomical classification. Then, we used the axonometric drawing to make the model more exact. Next, combining with some cases of pathological Riolan's arch and basing on hydrodynamic principle, we speculate the mechanism of collateral circulation. Finally, based on the retrospective study of high ligation cases and combined principles of fluid mechanics, we show how these shortcut anastomotic branches evolved into Riolan's arch. RESULTS: We report the classification of the ascending branch of LCA (which approaches the splenic flexure) and the left branch of MCA, display these shortcut anastomotic branches on a map, and establish their three-dimensional anatomical classification. We found that Riolan's arch is a shortcut pathway for the compensation of anastomotic branches, between MCA and LCA under pathological conditions, and that the formation mechanism of shortcut path accords with the principle of hydrodynamics. CONCLUSIONS: Our results show the mechanism of pathological Riolan's arch formation and provide new anatomic thinking for the battle between high and low ligation of IMA in colorectal cancer surgery.

Induction chemotherapy for unresectable Stage III non-small-cell lung cancer may improve survival of induction chemotherapy responders as predicted by elevated levels of carcinoembryonic antigen and cytokeratin fragment 19 and classification as stage N3 cancer.

AIMS: The aim of this study is to investigate patients with unresectable Stage III non-small-cell lung cancer (NSCLC) receiving radiotherapy with induction and concurrent pemetrexed or docetaxel plus cisplatin (PP/DP) chemotherapy and to identify the subgroup most likely to benefit from induction chemotherapy (IC). SUBJECTS AND METHODS: Patients with unresectable measurable Stage III NSCLC received two cycles of PP/DP IC followed by concurrent chemoradiotherapy at a dose of 60-66 Gy. STATISTICAL ANALYSIS USED: Cox regression analysis was performed to evaluate the prognostic factors for survival; logistic regression analysis was used to evaluate the predictors for response to IC, and the receiver operating characteristic curves were used to evaluate the independent factors predicting response. RESULTS: Eighty patients were included; the median survival time (MST) was 22.1 months. Partial response (PR) to IC was an independent prognostic factor for overall survival. For patients in the PR and stable disease groups, the MST was 36.7 and 19.5 months, respectively. The independent predictors of PR to IC included classification as stage N3 cancer, baseline carcinoembryonic antigen (CEA) levels >10 ng/ml, and cytokeratin fragment 19 (CYFRA21-1) levels >6 ng/ml. With each additional independent predictor, the likelihood of having have PR to IC increased. CONCLUSIONS: Radiotherapy with induction and concurrent PP/DP chemotherapy is feasible for patients with unresectable Stage III NSCLC. IC may improve the survival of IC responders, as predicted by elevated CEA and CYFRA21-1 levels and classification as stage N3 cancer. Additional randomized trials on IC may consider these predictors to tailor individualized treatments.

Hemoglobin, albumin, lymphocyte, and platelet score and neutrophil-to-lymphocyte ratio are novel significant prognostic factors for patients with small-cell lung cancer undergoing chemotherapy.

OBJECTIVE: The hemoglobin, albumin, lymphocyte, and platelet (HALP) score, neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR) are the important prognostic markers in some tumor types. This study aimed to evaluate the prognostic impact of pretreatment using HALP, NLR, and PLR for patients with small-cell lung cancer (SCLC), who were undergoing chemotherapy. MATERIALS AND METHODS: In this retrospective study, 335 patients with SCLC were included between 2016 and 2018. The cutoff values for HALP, NLR, and PLR were defined using X-tile software. Survival was analyzed by the Kaplan-Meier method, with differences analyzed through the log-rank test. The multivariate Cox proportional hazard model was used to evaluate the prognostic significance of HALP, NLR, and PLR for SCLC. RESULTS: The median follow-up period was 27.1 months (range: 0.5-46.2 months). Based on the Kaplan-Meier curve analysis, it was noticed that the low pretreatment HALP (≤18.6), high pretreatment NLR (>2.4), and high PLR (>191.6) were significantly associated with worse overall survival (OS) (P = 0.009, 0.001, and 0.033, respectively). Cox multivariate analysis demonstrated that low pretreatment HALP and high pretreatment NLR were the independent prognostic factors for worse OS (hazard ratio [HR] = 1.468, 95% confidence interval [CI]: 1.004-2.146, P = 0.047; HR = 0.722, 95% CI: 0.542-0.960, P = 0.025, respectively). CONCLUSION: HALP and NLR were the independent prognostic factors of OS for SCLC patients undergoing chemotherapy.

The oncogenic role of NKAP in the growth and invasion of colon cancer cells.

It has been reported that NF­κB activating protein (NKAP) is a transcriptional repressor of the Notch signaling pathway and is involved in the proliferation and survival of hematopoietic stem cells. In the present study, we aimed to investigate the effect of NKAP on the progression and metastasis of colon cancer. The results of immunohistochemical staining and western blot analysis showed that NKAP was upregulated in colon cancer tissues, and its expression was associated with colon cancer stages. CCK­8, colony formation, Transwell, and flow cytometry assays were used to demonstrate that NKAP knockdown significantly suppressed the proliferation and invasion of HCT116 and HT­29 cells, and also induced apoptosis and autophagy. By contrast, NKAP overexpression markedly promoted the proliferation and invasion of HCT­15 cells, and inhibited cell apoptosis and autophagy. Moreover, we observed that NKAP knockdown inhibited the epithelial­mesenchymal transition (EMT) process in HCT116 and HT­29 cells, while NKAP overexpression promoted EMT in HCT­15 cells. Furthermore, NKAP knockdown inhibited activation of the Akt/mTOR signaling pathway by downregulating the phosphorylation of Akt and mTOR, as well as their downstream proteins, whereas NKAP overexpression promoted the Akt/mTOR signaling pathway. Additionally, expression of P65 was downregulated by silencing of NKAP and upregulated by NKAP overexpression. These data suggest that NKAP functions as an oncogene in the growth and invasion of colon cancer in vitro.

Clinical observation of gene expression-guided chemoradiation therapy for nonsurgical esophageal squamous cell carcinoma patients: a retrospective analysis of 36 cases.

OBJECTIVE: To make an informed choice of chemotherapy drugs according to the oncogene mRNA expression and to explore whether it could increase the survival rate of patients. PATIENTS AND METHODS: The study retrospectively analyzed 36 cases of nonsurgical esophageal squamous cell carcinoma patients treated at the Center for Oncology of Shandong Provincial Hospital from December 1, 2010, to November 1, 2013. Intensity-modulated radiation therapy was used for the treatment with a conventional radiotherapy dose of 60-66 Gy. Chemotherapy started 1-5 weeks after radiation therapy. The selection of the chemotherapy drug was based on the mRNA expression levels of excision repair cross-complementation 1, thymidylate synthetase, ribonucleotide reductase M1, and ß-tubulin isotype III. The objective response rate, progression-free survival, and overall survival were observed. RESULTS: The reason for poor prognosis of patients with high expression of excision repair cross-complementation 1 was unknown. No correlation was observed between patient survival and expression of thymidylate synthetase, ribonucleotide reductase M1, and ß-tubulin isotype III. Complete response, partial response, stable disease, and progressive disease were observed in 25, five, three, and three patients, respectively. The objective response rate was 83.3%. The 1-year, 2-year, and 3-year progression-free survival rates were 79.8%, 58.9%, and 54.4%, respectively. The 1-year, 2-year, and 3-year overall survival rates were 83.3%, 68.1%, and 58.4%, respectively. CONCLUSION: Selecting the chemotherapy drug according to the oncogene expression, combined with radiation therapy, could increase the 3-year survival rate in nonsurgical esophageal squamous cell carcinoma patients. Such conclusion needs to be further confirmed using a larger sample size.