TYMS Knockdown Suppresses Cells Proliferation, Promotes Ferroptosis via Inhibits PI3K/Akt/mTOR Signaling Pathway Activation in Triple Negative Breast Cancer.

Triple-negative breast cancer (TNBC) is characterized by a grim prognosis and numerous challenges. The objective of our study was to examine the role of thymidylate synthase (TYMS) in TNBC and its impact on ferroptosis. The expression of TYMS was analyzed in databases, along with its prognostic correlation. TYMS positive expression was identified through immunohistochemistry (IHC), while real-time quantitative PCR (qRTPCR) was employed to measure TYMS mRNA levels in various cell lines. Western blotting was utilized to assess protein expression. Cell proliferation, mobility, apoptosis, and reactive oxygen species (ROS) levels were evaluated using CCK8, wound scratch healing assay, transwell assay, and flow cytometry, respectively. Additionally, a tumor xenograft model was established in BALB/c nude mice for further investigation. Tumor volume and weight were measured, and histopathological analysis using hematoxylin and eosin (H&E) staining was conducted to assess tumor tissue changes. IHC staining was employed to detect the expression of Ki67 in tumor tissues. High expression of TYMS was observed in TNBC and was found to be correlated with poor prognosis in patients. Among various cell lines, TYMS expression was highest in BT549 cells. Knockdown of TYMS resulted in suppression of cell proliferation and mobility, as well as promotion of apoptosis. Furthermore, knockdown of TYMS led to increased accumulation of ROS and Fe2+ levels, along with upregulation of ACLS4 expression and downregulation of glutathione peroxidase 4 (GPX4) expression. In vivo studies showed that knockdown of TYMS inhibited tumor growth. Additionally, knockdown of TYMS was associated with inhibition of mTOR, p-PI3K, and p-Akt expression. Our research showed that the knockdown of TYMS suppressed the TNBC progression by inhibited cells proliferation via ferroptosis. Its underlying mechanism is related to the PI3K /Akt pathway. Our study provides a novel sight for the suppression effect of TYMS on TNBC.

The impact of body mass index on operative time in transoral endoscopic thyroidectomy vestibular approach for thyroid cancer.

BACKGROUND: The impact of body mass index (BMI) on operative time in transoral endoscopic thyroidectomy vestibular approach (TOETVA) for thyroid cancer is still a subject of debate. This study assessed the impact of BMI on operative time and postoperative complications in patients undergoing TOETVA. METHODS: The study has been conducted to compare the outcomes of TOETVA in patients with high BMI (≥25) and those with normal BMI (<25). Postoperative outcomes, including operative time, blood lost, recurrent laryngeal nerve (RLN) palsy, hypocalcemia and postoperative pain score, were evaluated. RESULTS: A total of 62 patients who underwent TOETVA were included in the study. The high BMI group consisted of 39 patients, while the normal BMI group included 23 patients. No significant differences were observed between the two groups regarding operative time, blood loss, postoperative pain score, and postoperative complications such as recurrent laryngeal nerve (RLN) palsy and hypocalcemia. CONCLUSIONS: BMI was not significantly associated with operative time and postoperative complications in patients undergoing TOETVA, indicating its safety and feasibility for elevated BMI patients.

Prediction and screening of circRNA in triple-negative breast cancer.

OBJECTIVE: The purpose of this investigation was to study the expression profile and potential function of circular RNA (circRNA) and long noncoding RNA (lncRNA) in triple-negative breast cancer (TNBC). METHODS: RNA sequencing technology was used to detect differentially expressed circRNAs and lncRNAs between TNBC tissues and the adjacent tissue. The potential functions of these different RNAs were analyzed by GO and KEGG enrichment analysis by bioinformatics tools. We also selected and analyzed these key circRNAs and lncRNAs to verify their important functions in TNBC. RESULTS: A total of 139 differentially expressed circRNAs and 1001 lncRNAs were obtained. The co-expression analysis showed that the hub lncRNAs (OIP5-AS1, DRAIC) were associated with several tumors and mainly enriched in tumor metastasis. We also screened 5 circRNA-hosting genes (NTRK2, FNTA, BAPGEF2, MGST2, ADH1B) that were associated with the brain-derived neurotrophic factor (BDNF) receptor signaling pathway and cerebral cortex development, as well as AMPK and TGF-ß signaling pathway. CONCLUSION: We identified a large number of differentially expressed circRNAs and lncRNAs, which provide useful insight in understanding TNBC carcinogenesis.

[Clinical feasibility of imaging with indocyanine green combined with methylene blue for sentinel lymph node identification in papillary thyroid microcarcinoma].

Objective:To explore the value of indocyanine green（ICG） combined with methylene blue in the identification of sentinel lymph nodes（SLNs） in patients diagnosed with papillary thyroid microcarcinoma（PTMC）. Methods:Ninety patients were enrolled and were randomized into group A and group B with 45 patients in each group. ICG combined with methylene blue were injected into the thyroid in group A, and only methylene blue were injected into thyroid in group B. Blue-stained or fluorescent nodes observed using near-infrared fluorescence imaging systems were defined as SLNs. After SLNs were removed, central lymph nodes（CLNs） dissection was completed in both groups. The pathological data and postoperative outcomes were compared between two groups. Results:There were significantly more SLNs（2.93/2.17） and CLNs（4.51/3.89） were dissected in group A than in group B（P<0.05）. There were no significant differences in sensitivity, accuracy and the false-negative rate according the SLNs in two groups（P>0.05）, but group A has higher sensitivity and accuracy rates, and lower false-negative rate. There were no significant differences in the amount of blood loss, the amount of lymphatic drainage, and incidence of hoarseness and lymphatic leakage in two groups（P>0.05）. In group A, the operating time was longer, and the rate of hypoparathyroidism was lower（P<0.05）. Conclusion:Sentinel lymph nodes biopsy using ICG combined with methylene blue is feasible and safe for SLNs identification in PTMC patients. It is also clinically significant for the parathyroid gland protection.

LncRNA ZFAS1/miR-589 regulates the PTEN/PI3K/AKT signal pathway in the proliferation, invasion and migration of breast cancer cells.

Breast cancer (BC) is a common clinical disease and the second leading cause of cancer death in women. Long noncoding RNA (lncRNA) and microRNA (miRNA) are reported to be involved in the development of BC. The present study aimed to investigate whether LncRNA ZFAS1 could regulate the proliferation, invasion and migration of breast cancer cells by targeting miR-589 through PTEN/PI3K/AKT signal pathway. The expression of ZFAS1 and miR-589 in BC cells and transfection effects were determined by RT-qPCR analysis. The abilities of proliferation, colony formation, invasion and migration of breast cancer cells were analyzed by CCK-8 assay, colony formation assay, transwell assay and wound healing assay respectively. The expression of MMP2, MMP9, Bcl2, Bax, cleaved caspase3, PTEN, p-PI3K, p-AKT, PI3K and AKT was detected by Western blot. The flow cytometry analysis was used to detect cell apoptosis. As a result, ZFAS1 expression was increased and miR-589 expression was decreased in BC cells. And, miR-589 was demonstrated to be a target of ZFAS1. ZFAS1 overexpression could inhibit the proliferation, colony formation, invasion and migration of BC cells while miR-589 overexpression could reverse the changes. In addition, ZFAS1 overexpression suppressed the expression of PI3K/AKT signal pathway by activating the PTEN expression while miR-589 overexpression could reverse the changes. Moreover, PTEN is one of the gene targets of miR-589. In conclusion, this study indicated that ZFAS1 inhibited the proliferation, invasion and migration of breast cancer cells by targeting miR-589 through regulating the PTEN/PI3K/AKT signal pathway.

Connexin 32 downregulation is critical for chemoresistance in oxaliplatin-resistant HCC cells associated with EMT.

Background: Oxaliplatin (OXA)-based chemotherapy is critical in the management of advanced hepatocellular carcinoma (HCC); however, acquired drug resistance has largely restricted its clinical efficacy. This study aims to explore the key mechanisms and regulatory factors determining chemosensitivity in HCC. Methods: We developed OXA-resistant (OR) HCC cells and used multiple methods, including real-time RT-PCR, Western blot, immunofluorescence, transwell invasion assay, wound-healing assay, MTT assay, gene transfection, and immunohistochemistry to achieve our goals. Results: We found that OR HCC cells showed a typical epithelial-mesenchymal transition (EMT) phenotype. Meanwhile, the expression of Cx32, a major member of the liver connexin (Cx) family, was lowly expressed in OR HCC cells. Downregulation of Cx32 in parental HCC cells led to EMT induction and thereby reduced OXA cytotoxicity, while Cx32 upregulation in OR HCC cells could reverse the EMT phenotype and partially restore chemosensitivity to OXA. Finally, in human HCC tissue samples, Cx32 was positively correlated with the expression of the EMT marker E-cadherin and negatively correlated with the expression of Vimentin. Conclusion: Our findings demonstrated that downregulation of Cx32 may be an important determinant for HCC cells to acquire EMT-related acquired drug resistance to OXA, and targeting Cx32 could be a novel strategy to overcome OXA resistance in HCC.

Evaluation of the clinical efficacy of preserving the left colic artery in laparoscopic resection for rectal cancer: A meta-analysis.

The aim of the present meta-analysis compared left colic artery (LCA) preservation with non-preservation in laparoscopic resection of rectal cancer in terms of feasibility, efficacy and safety. The PubMed, Ovid, Embase, Web of Science, CBM, CNKI, VIP and WanFang Data databases were searched prior to June 2017 for studies comparing LCA preservation and non-preservation in laparoscopic resection for rectal cancer. Two researchers screened the literature independently, extracted the data and evaluated the risk of bias. The study was performed using RevMan 5.3 software for meta-analysis. A total of 10 studies comparing LCA preservation and non-preservation in laparoscopic resection for rectal cancer were selected for this meta-analysis, with a combined study population of 1,471 patients. The results of the meta-analysis demonstrated that, when comparing LCA preservation with non-preservation in laparoscopic resection for rectal cancer, there were significant differences between the two groups in terms of operative time (P<0.01), estimated blood loss (P<0.01), percentage of neostomy (P<0.01), the number of retrieved lymph nodes (P<0.01), time to first postoperative exhaust (P<0.01) and amount of anastomotic leakage (P<0.01). However, there were no significant differences in postoperative hospital stay (P=0.28), incidence of recurrence (P=0.73) and incidence of metastasis (P=0.52). Therefore, compared with LCA non-preservation, patients in whom the LCA was preserved during laparoscopic resection for rectal cancer had a better prognosis. However, there was no difference in recurrence or metastasis between the two groups. Although the operative time and estimated blood loss were increased with LCA preservation, these may be reduced with improving proficiency of the operating surgeons. The conclusions of the present study require verification by larger samples and high-quality randomized controlled trials.

Effects and mechanism of flavonoids from Astragalus complanatus on breast cancer growth.

Flavonoids from Astragalus complanatus R.Br (FAC) had anticancer effects on many tumor cells. The current study was performed to evaluate the effects of FAC on human breast cell proliferation, apoptosis, and metastasis, as well as their active mechanism. Cell viability and growth were detected using the cell counting kit (CCK)-8 assay in vitro. Assay of FAC on induced breast cancer mortality was counted as survival time of nude mice after breast cell line inoculation. The effect of FAC on cell invasion was investigated by an optimization assay that contains a 96-well Boyden chamber with wells precoated with BME at three different concentrations. The mechanism of its action on apoptosis and metastasis was determined by related gene detection. Proliferation of three breast cell lines was inhibited with FAC treatment in a dose-dependent manner. Survival time of nude mice with breast cancer cell inoculation also was prolonged with increasing FAC dose. Metastasis in FAC-treated breast cells was also significantly inhibited. Real-time polymerase chain reaction (PCR) assay demonstrated that apoptosis-related BCL-2 and caspase-9 gene expression was consistent with their phenotype change. Metastasis-related FAK and BRCA1 gene expression was inversely related to FAC treatment. Western blot analysis indicated that BCL-2 and FAK proteins were reduced, whereas caspase-9 and BRCA1 proteins were increased with a higher dose of FAC treatment. These data suggested that FAC has an important role in breast cancer growth and metastasis suppression in vitro and in vivo. Its active mechanism involves promoting programmed cancer cell death and regulates metastasis-related gene expression.

AZD-4547 exerts potent cytostatic and cytotoxic activities against fibroblast growth factor receptor (FGFR)-expressing colorectal cancer cells.

Colorectal cancer (CRC) causes significant mortalities worldwide. Fibroblast growth factor (FGF) receptor (FGFR) signaling is frequently dysregulated and/or constitutively activated in CRCs, contributing to cancer carcinogenesis and progression. Here, we studied the activity of AZD-4547, a novel and potent FGFR kinase inhibitor, on CRC cells. AZD-4547 inhibited CRC cell growth in vitro, and its activity correlated with the FGFR-1/2 expression level. AZD-4547 was cytotoxic and pro-apoptotic in FGFR-1/2-expressed CRC cell lines (NCI-H716 and HCT-116), but not in FGFR-1/2 null HT-29 cells. Further, AZD-4547 inhibited cell cycle progression and attenuated the activation of FGFR1-FGFR substrate 2 (FRS-2), ERK/mitogen-activated protein kinase (MAPK), and AKT/mammalian target of rapamycin (AKT/mTOR) signalings in NCI-H716 and HCT-116 cells. In vivo, AZD-4547 oral administration at effective doses inhibited NCI-H716 (high FGFR-1/2 expression) xenograft growth in nude mice. Phosphorylation of FGFR-1, AKT, and ERK1/2 in xenograft specimens was also inhibited by AZD-4547 administration. Thus, our preclinical studies strongly support possible clinical investigations of AZD-4547 for the treatment of CRCs harboring deregulated FGFR signalings.

[Efficacy of modified radical operation of preserving nipple-areolar complex on early stage breast cancer].

BACKGROUND AND OBJECTIVE: Modified radical mastectomy of preserving nipple-areolar complex (NAC) is an important surgical therapy for stage I-IIa breast cancer, but the oncological risk is controversial. This study was to compare the efficacy of NAC-preserving modified radical operation and conventional modified radical operation on early stage breast cancer. METHODS: The patients who received NAC-preserving modified radical operation (42 patients) or conventional modified radical operation (84 patients) from January 1998 to December 2003 in the First Affiliated Hospital of Bengbu Medical College were matched with a ratio of 1:2 by age at diagnosis, axillary lymph node status, sexual hormone receptor status, tumor size and Her-2/neu expression for retrospective analysis. The loco-regional recurrence, distant metastasis, 5-year disease-free survival (DFS) and 5-year overall survival (OS) between the two groups were compared. RESULTS: Median follow-up time was 48 months in NAC-preserving operation group and 44 months in conventional operation group. The 5-year occurrence rate of loco-regional recurrence was 2.44% in NAC-preserving operation group and 3.21% in conventional operation group (P=0.771). The 5-year occurrence rate of distant metastasis was 5.64% in NAC-preserving operation group and 4.30% in conventional operation group (P=0.654). The 5-year OS rates were 96.00% in NAC-preserving operation group and 98.18% in conventional operation group (P=0.694). The 5-year DFS rates were 91.67% in NAC-preserving operation group and 92.26% in conventional operation group (P=0.597). CONCLUSION: Modified NAC-preserving radical operation results in the same effect on early stage breast cancer as conventional modified radical operation based on careful consideration of the indications, and results in better cosmetic appearance after restitution and better quality of life.

[Clinicopathologic analysis of the nipple-areolar complex occult involvement in early stage breast carcinoma].

OBJECTIVE: The aim of this study was to investigate the incidence of nipple-areola complex (NAG) involvement in stage I - II a breast cancer patients who underwent skin-sparing mastectomy and to determine the associated risk factors, to provide a theoretical basis for modified radical mastectomy preserving NAC and breast reconstruction in early stage breast cancer patients. METHODS: A total of 68 women with primary breast cancer were included in this study. The following associated risk factors of NAC involvement were assessed and compared with those of non-involvement: the distance from the tumor site to the edge of areola (D), axillary lymph node status, over-expression of HER-2/neu, location of tumor, TNM stage, abnormal nipple (nipple indentation, erosion, discharge), tumor size, age, histological type, as well as status of estrogen receptor (ER) and progesterone receptor (PR), by Chi-square test. RESULTS: The positive rate of NAG involvement was 13.2%. It decreased with an increase in the distance from the tumor site to the edge of the areola (D) (chi2 = 10.68, P <0.01)), and higher incidence of NAG involvement was found in patients with axillary lymph node metastasis (chi2 = 14. 61, P < 0.01) and over-expression of HER-2/neu (chi2 =6.83, P <0.01). Location of tumor (P <0.01), TNM stage (chi2 =3.85, P <0.05), abnormal nipple (chi2 = 11.65, P<0.01), and tumor size (chi2 =4.13, P <0.05) also had influence on the NAG involvement. No significant correlation between NAC involvement and age (P > 0.05)), histological type (chi2 = 0.07, P > 0.05)), as well as status of estrogen receptor (ER) (chi2 = 0.06, P > 0.05) and progesterone receptor (PR) (chi2 = 0.04, P > 0.05) was found. Most of the NAG involvement was caused by ductal infiltration. CONCLUSION: In the stage I - II a breast cancer patients, location of tumor, TNM stage, the distance from the tumor site to the edge of areola (D), abnormal nipple, over-expression of HER-2 and metastases in axillary lymph nodes are the primary influential factors of NAG involvement.