Discussion on the Treatment Strategy for Stage â ¡A1 Cervical Cancer (FIGO 2018).

Objective: This study aimed to explore the best treatment strategy for International Federation of Gynecology and Obstetrics (FIGO) 2018 stage IIA1 cervical cancer patients by comparing the survival outcomes of two treatment methods: abdominal radical hysterectomy (ARH) with standard postoperative therapy and radio-chemotherapy (R-CT). Methods: Patients with FIGO2018 stage IIA1 cervical cancer who underwent ARH or received R-CT were screened from the clinical diagnosis and treatment for cervical cancer in China (Four C) database. The recurrence cases between the two groups were analyzed. The 5-year overall survival (OS) and disease-free survival (DFS) of patients diagnosed with stage IIA1 cervical cancer in 47 hospitals in mainland China between 2004 and 2018 were compared by using propensity score matching (PSM). Results: A total of 724 patients met the inclusion criteria. In the total study population, The R-CT group had higher recurrence (22.8% for the R-CT group and 11.2% for the ARH group, P<0.001) rates compared to the ARH group.The 5-year OS and DFS of the ARH group (n=658) were significantly higher than those of the R-CT group (n=66) (OS: 85.9% vs. 71.2%, P=0.009; DFS: 79.2%vs. 70.5%, P=0.027). R-CT was associated with worse 5-year OS (HR=3.19, 95% CI: 1.592-6.956, P=0.001) and DFS (HR=2.089, 95% CI: 1.194-3.656, P=0.01). After 1:2 PSM, the 5-year OS and DFS of the ARH group (n=126) were significantly higher than those of the R-CT group (n=64) (OS:88.9% vs. 70.1%, P=0.04; DFS:82.8% vs. 69.8%, P=0.019). R-CT was still associated with worse 5-year OS (HR=2.391, 95% CI: 1.051-5.633, P=0.046) and DFS (HR=2.6, 95% CI: 1.25-5.409, P=0.011). Conclusion: Our study demonstrated that for stage FIGO2018 stage IIA1 cervical cancer patients, ARH offers better oncological outcomes than R-CT.

Mitochondrial fission regulator 2 (MTFR2) promotes growth, migration, invasion and tumour progression in breast cancer cells.

INTRODUCTION: Mitochondrial fission regulator 2 (MTFR2) belongs to the MTFR family, and 2 isoforms of MTFR2 are produced by alternative splicing. The role of MTFR2 in breast cancer (BC) remains unknown. RESULTS: MTFR2 was upregulated in BC tissues and was strongly associated with tumor characteristics. Moreover, Kaplan-Meier and Cox proportional hazards analyses indicated that high MTFR2 expression was related to poor overall survival. In addition, the capacity for migration and invasion decreased in two BC cell lines after knockdown of MTFR2. The epithelial-mesenchymal transition pathway was inhibited in MTFR2-silenced cells. MTFR2 can switch glucose metabolism from OXPHS to glycolysis in a HIF1α- and HIF2α-dependent manner. CONCLUSION: Taken together, our results indicate that increased expression of MTFR2 is associated with tumour progression in breast cancer cells through switching glucose metabolism from OXPHS to glycolysis in a HIF1α- and HIF2α-dependent manner. MATERIALS AND METHODS: We obtained data from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) to analyse MTFR2 expression in BC. The prognostic value of MTFR2 expression was assessed using the Kaplan-Meier method. The biological influence of MTFR2 on BC cell lines was studied using proliferation, Transwell migration, invasion and mitochondrial function assays.

Long noncoding RNA LINC00511 promotes cell growth and invasion in triple-negative breast cancer by interacting with Snail.

Purpose: Aberrant long noncoding RNA expression has been frequently reported in cancer research, including in triple-negative breast cancer (TNBC). The aim of the present study was to investigate the involvement of LINC00511 in the progression and prognosis of TNBC. Materials and methods: The expression level of LINC00511 was examined by RT-PCR in TNBC tissues and in cell lines. MTT and colony formation assays were used to examine the cell growth ability. A Boyden assay was used to examine the cell invasion ability. RNA pull-down and RNA immunoprecipitation (RIP) assays were used to examine the proteins that interacted with LINC00511. Results: We demonstrated that the LINC00511 expression level was elevated in TNBC tissues when compared with that in normal breast tissues. The downregulation of LINC00511 decreased TNBC cell growth and invasion compared to those of the controls. To explore the molecular mechanisms underlying the biological activity of LINC00511, we identified proteins that bound to LINC00511 with RNA pull-down experiments. We showed that LINC00511 binds to the ß-transducin repeat containing (BTRC) E3 ubiquitin protein. Mechanistically, LINC00511 maintained the stability of Snail by impeding its ubiquitination and degradation by the BTRC E3 ubiquitin protein. Conclusion: Our data suggested that LINC00511 might serve as a novel molecular target for the treatment of TNBC.

Expression status of cytoskeleton regulatory protein Mena as a prognostic marker for human thyroid carcinoma.

OBJECTIVE: In recent years, Mena (Mammalian Enabled) has been reported to be highly expressed in malignant tumors. However, data on the expression pattern and clinical relevance of Mena in thyroid carcinoma are unclear. The purpose of this study was to investigate the expression of Mena and its prognostic significance in human thyroid carcinoma. MATERIALS AND METHODS: Mena expression at the mRNA level was examined by real-time quantitative polymerase chain reaction (RT-PCR) in 8 paired thyroid carcinoma and adjacent normal tissues. Mena protein expression in clinical samples was analyzed in paraffin-embedded papillary thyroid carcinoma samples and normal thyroid tissues by immunohistochemistry (IHC). Statistical analyses were also performed to evaluate the clinicopathological significance of Mena expression. RESULTS: The results show that expression of Mena mRNA is higher in thyroid carcinoma than in adjacent normal tissues in 8 paired samples. In paraffin-embedded tissue samples, the expression of Mena was higher in papillary thyroid carcinoma than normal thyroid tissues. Compared with normal thyroid tissues, overexpression of Mena was detected in 47.11% (57/121) of papillary thyroid carcinoma patients. Overexpression of Mena was significantly associated with T Stage (P = 0.007), capsular invasion (P = 0.015), lymph node metastasis (P = 0.000), and clinical stage (P = 0.029). CONCLUSION: Mena is up-regulated in thyroid carcinoma and is associated with expression of T Stage, lymph node metastasis, clinical stage and disease-free survival. Mena may serve as a prognostic indicator for patients with thyroid carcinoma.

A new method of subtotal thyroidectomy for Graves' disease leaving a unilateral remnant based on the upper pole.

BACKGROUND: The aim of this prospective randomized study was to evaluate the feasibility of subtotal thyroidectomy with leaving a unilateral remnant based on the upper pole. METHODS: Patients who underwent the subtotal thyroidectomy and isthmusectomy leaving either a unilateral remnant based on the upper pole (Group I, 79 patients) or the bilateral dorsal thyroid tissue remained (Group II, 89 patients) were compared in operation time, blood loss, recurrence, and postoperative complications. RESULTS: Among 168 patients analyzed, the operation time remained similar, but the blood loss, the reoperation time, and recurrence in Group I were much less than Group II. In addition, no postoperative hemorrhage occurred in Group I. Two patients (2.28%) in Group II underwent recurrent laryngeal nerve damages. Four patients (5.06%) in Group I and 3 patients (3.37%) in Group II experienced transient hypocalcemia. Recurrence only occurred in Group II. CONCLUSION: In terms of blood loss, reoperation time, postoperative complication, and the recurrence, subtotal thyroidectomy with recurrent laryngeal nerves identification and the unilateral superior pole remnant of the gland provides a better outcome than subtotal thyroidectomy with bilateral dorsal thyroid tissue remnant.

Expression of Gab1 Is Associated with Poor Prognosis of Patients with Epithelial Ovarian Cancer.

Growth factor receptor-bound protein-2 (Grb2) can act as the scaffold protein recruiting other molecules to the stimulated receptors. Grb2-associated binding protein 1 (Gab1) is involved in cell proliferation, and its expression may enhance the carcinogenesis and cancer progression. However, the function of Gab1 remains to be investigated. Epithelial ovarian cancer (EOC) is the most lethal malignancy in the female reproductive system with increasing incidence and unsatisfied overall survival (OS). We investigated the expression of Gab1 in EOC tissues and the correlations between Gab1 expression and the clinicopathological characteristics of patients with EOC using Spearman rank test. The staining results were evaluated based on both the percentage of Gab1-positive tumor cells and the staining intensity for Gab1 expression. Kaplan-Meier survival analysis and Cox proportional hazards analysis were used to compare the postoperative OS between EOC patients with high Gab1 expression and those with low Gab1 expression. The high expression of Gab1 was positively correlated with advanced FIGO stage and lymph node metastasis of EOC. Univariate analysis showed that advanced FIGO stage, pathological grade, lymph node metastasis or Gab1 expression were associated with poor OS. Moreover, multivariate analysis revealed that Gab1 expression could be an independent prognostic factor for the poor OS of EOC patients (P = 0.042). We propose that Gab1 expression is correlated with poor prognosis of EOC patients and may act as an independent prognostic indicator.

Hsa-miR-1 suppresses breast cancer development by down-regulating K-ras and long non-coding RNA MALAT1.

MicroRNAs exert their functions by mainly regulating coding genes or long non-coding RNA expression. In the present study, we reported that hsa-miR-1 was down-regulated in breast cancer tissues. Restoration of miR-1 in breast cancer cells inhibited proliferation, motility and increased apoptosis in vitro. MiR-1 functioned as a tumor suppressor by targeting K-RAS and MALAT1. In addition, the effects of up-regulation of miR-1 were similar to that of silencing K-RAS and MALAT1 in breast cancer cells. In vivo study indicated that restoration of miR-1 inhibited tumor growth and metastasis. Patients with low miR-1 expression had poorer overall survival time than those with high miR-1 expression. Our findings emphasized the potential role of miR-1 as tumor suppressive miRNA in breast cancer.

Tumor suppressor miR-145 reverses drug resistance by directly targeting DNA damage-related gene RAD18 in colorectal cancer.

Colorectal cancer (CRC) is one of the most common cancers worldwide. Although chemotherapy is used as a palliative treatment, ultimately, nearly all patients develop drug resistance. Therefore, the cell-inherent DNA repair pathway must reverse the DNA-damaging effect of cytotoxic drugs that mediates therapeutic resistance to chemotherapy. RAD18, a DNA damage-activated E3 ubiquitin ligase, is known to play a critical role in DNA damage repair in cancer cells. Here, we show that RAD18 is highly expressed in human 5-fluorouracil (5-FU)-resistant cancer cells after 5-FU treatment. In addition, RAD18 increases in CRC cells could induce DNA damage repair, suggesting that RAD18 might be a possible target for overcoming drug resistance. Moreover, the expression of tumor suppressor microRNA-145 (miR-145) was negatively correlated with RAD18 expression in CRC tissues of 140 patients. Using luciferase reporters carrying the 3'-untranslated region of RAD18 combined with Western blotting, we identified RAD18 as a direct target of miR-145. Also of interest, suppression of RAD18 by miR-145 enhanced DNA damage in CRC cells after 5-FU treatment. Finally, the 5-FU-resistant cancer cells could be selectively ablated by treatment with miR-145. Taken together, these results suggest that miR-145 can act as an RAD18 inhibitor and contribute as an important factor in reversing drug resistance after chemotherapy.

[Establishment of a NOD/SCID mouse model with human immune reconstitution bearing human triple-negative breast cancer].

OBJECTIVE: To establish a NOD/SCID mouse model with human immune reconstitution and observe its immune response to human triple-negative breast cancer xenograft. METHODS: Twenty-four NOD/SCID mice without immune leakage were subjected to cyclophosphamide (CTX) treatment 3 days prior to immune reconstitution with human peripheral blood mononuclear cell (PBMC) injection and subcutaneous transplantation of human triple-negative breast cancer MDA-MB-231 cells, CTX treatment and PBMC injection without tumor cell transplantation, MDA-MB-231 cell transplantation only, or no treatments. The tumor growth and immune responses of the mice were observed at regular intervals. RESULTS: Compared with the tumor-bearing mice, the tumor-bearing mice with immune reconstitution showed prolonged incubation period of tumor formation, slower tumor growth rate and increased survival rate. Human IgG and CD3(+) T cells were detected in the peripheral blood of the mice 1 week after human PBMC injection. The percentage of CD3(+) T cells in the spleen cells was 55.3% at 9 weeks in tumor-bearing mice with immune reconstitution and 52.7% in tumor-bearing mice without immune reconstitution. The spleen index of the tumor-bearing mice with immune reconstitution was much higher than that in mice with only immune reconstitution and the control mice (9.64 vs 3.82∓0.31 and 1.51∓0.14 mg/g). CONCLUSION: A stable NOD/SCID mouse model with immune reconstitution has been established successfully, which shows immune responses to triple-negative breast cancer xenografts and allows studies of immunological therapy study of triple-negative breast cancer.

Preparation of triple-negative breast cancer vaccine through electrofusion with day-3 dendritic cells.

Dendritic cells (DCs) are professional antigen-presenting cells (APCs) in human immune system. DC-based tumor vaccine has met with some success in specific malignancies, inclusive of breast cancer. In this study, we electrofused MDA-MB-231 breast cancer cell line with day-3 DCs derived from peripheral blood monocytes, and explored the biological characteristics of fusion vaccine and its anti-tumor effects in vitro. Day-3 mature DCs were generated from day-2 immature DCs by adding cocktails composed of TNF-α, IL-1ß, IL-6 and PEG2. Day-3 mature DCs were identified and electofused with breast cancer cells to generate fusion vaccine. Phenotype of fusion cells were identified by fluorescence microscope and flow cytometer. The fusion vaccine was evaluated for T cell proliferation, secretion of IL-12 and IFN-γ, and induction of tumor-specific CTL response. Despite differences in morphology, day-3 and day-7 DC expressed similar surface markers. The secretion of IL-12 and IFN-γ in fusion vaccine group was much higher than that in the control group. Compared with control group, DC-tumor fusion vaccine could better stimulate the proliferation of allogeneic T lymphocytes and kill more breast cancer cells (MDA-MB-231) in vitro. Day-3 DCs had the same function as the day-7 DCs, but with a shorter culture period. Our findings suggested that day-3 DCs fused with whole apoptotic breast cancer cells could elicit effective specific antitumor T cell responses in vitro and may be developed into a prospective candidate for adoptivet immunotherapy.

[Application of in vivo pelvis and abdominal-pelvic arterial network digital three-dimensional model for uterine artery embolization in surgical approach planning].

OBJECTIVE: To investigate the construction and significance of in vivo pelvis and abdominal-pelvic arterial vascular network in digital three-dimensional (3D) model for uterine artery embolization (UAE) in the application of surgical approach planning based on computerized tomographic angiography(CTA). METHODS: A series of digital imaging and communications in medicine 3.0 (Dicom 3.0) were obtained from a woman with myoma of uterus by CTA scanning. Then the software Mimics Version 10.01 was used to construct the pelvic and the arterial vascular network 3D model. RESULTS: The digital model could clearly display the abdominal aorta, bilateral common iliac arteries, left and right external iliac artery, internal iliac artery and its branches around the stage; the right uterine artery emitted in the inferior gluteal artery opening below 15.91 mm, the left uterine artery in inferior gluteal artery below the opening of 15.21 mm, the adjacent artery of internal pudendal artery. At the same time, the artery length and angle of bifurcation were accurately measured. The bifurcation angle of abdominal aorta in fifth lumbar vertebral body edge, was 66.58°, the bifurcation angle of right internal iliac artery was 46.23°, the length of right common iliac artery was 51.43 mm, the bifurcation angle between left and external internal iliac artery was 36.45°, the length of left common iliac artery and 67.50 mm. According to the preoperative approach planning, guided wire across the inferior gluteal artery to the lower 15.00 mm and rotating digital subtraction angiography (DSA) tube, could clearly display the uterine artery opening outward by rotating guide wire, which was relatively easy to enter the uterine artery. CONCLUSION: The female pelvic arterial network model in vivo could be successfully constructed by using Mimics Version 10.01 software with database collected through CTA, which may contribute to the materialization of digital models and be used for preoperative surgical simulator.

Triptolide-induced in vitro and in vivo cytotoxicity in human breast cancer stem cells and primary breast cancer cells.

We investigated the potential efficacy of the Chinese herbal extract triptolide for the treatment of human breast cancer by measuring the triptolide-induced cytotoxicity in cultures of human primary breast cancer cells (BCCs) and breast cancer stem cells (BCSCs) in vitro and in vivo. Human BCCs and BCSCs from invasive ductal carcinoma samples were cultured and treated with 0.1, 0.5 or 1.0 µM triptolide. Cell death and apoptosis were measured after 24, 48 and 72 h of treatment. Mammospheres were found to be highly tumorigenic when implanted subcutaneously in nude BALB/c mice. Triptolide was cytotoxic against both human primary BCCs and BCSCs in vitro (P<0.05), but the cytotoxicity was stronger against the BCCs. In response to 1 µM triptolide for 72 h, the apoptotic rates were approximately 60% for BCCs and 30% for BCSCs. The BCSCs exhibited a high formation rate of tumors when implanted subcutaneously in nude BALB/c mice. Triptolide treatment in vivo significantly inhibited tumor growth compared with mock treatment. In conclusion, the cytotoxicity of triptolide against BCCs and BCSCs in vitro and in vivo suggests that this natural diterpenoid triepoxide compound may have clinical applications for the suppression of breast tumor growth.

miR-124 radiosensitizes human glioma cells by targeting CDK4.

The aberrant expression of cyclin-dependent kinase-4 (CDK4) has previously been observed in human brain glioma. Furthermore, it is observed that up-regulation of CDK4 is associated with therapy resistance and relapse. However, the mechanisms behind these phenomena remain unclear. Here, we demonstrated that elevated CDK4 expression is correlated with poor prognosis in glioma after radiotherapy and that CDK4 knockdown conferred radiosensitivity in glioma cell lines. CDK4 was identified as potential downstream target of miR-124 through bioinformatics analysis and dual-firefly luciferase reporter assay. Furthermore, restoration of miR-124 could confer radiosensitivity. Cell differentiation agent-2 (CDA-2) mimicked the effect of miR-124 restoration and CDK4 knockdown, and sensitized xenografts to radiation in an animal model. Our findings demonstrated for the first time that CDK4 was a downstream target of miR-124 and that CDA-2 could radiosensitize Glioblastoma multiforme cells through the MiR-124-CDK4 axis.

[Antitumor effect of a triple negative breast cancer-dendritic cell fusion vaccine].

OBJECTIVE: To test the antitumor effect of a human triple-negative breast cancer cell-dendritic cell (DC) fusion vaccine. METHODS: DCs were isolated from fresh peripheral blood of healthy donors. The fusion vaccine was prepared by fusing the DCs and MDA-MB-231 cells via electrofusion. The morphology of the vaccine was identified under inverted fluorescence microscope and the phenotypes were analyzed with flow cytometry. The production of interleukin-12 (IL-12) and interferon-γ (IFN-γ) by the fusion cells was assessed using ELISA. A CCK-8 kit was used to examine the effect of the vaccine in stimulating the proliferation and cytotoxicity of autologous T lymphocytes. RESULTS: The DCs isolated from peripheral blood mononuclear cells highly expressed CD83, CD86, CD11c and HLA-DR on the cell surface. The fusion cells were irregular in shape and coexpressed the phenotypes of DCs and MDA-MB-231 cells. The fusion cells possessed a strong ability to stimulate the proliferation of T lymphocytes in vitro. Compared with the control group, the fusion vaccine showed a stronger antitumor effect against the breast cancer cells. CONCLUSION: The triple-negative breast cancer-DC fusion vaccine prepared by electrofusion can stimulate the proliferation of T lymphocytes and induces strong cytotoxicity of the T cells against breast cancer cells.