“三阴”乳腺癌新辅助化疗前后BCSG1基因状况研究

目的探讨乳腺癌特异基因(BCSG1)在"三阴"性乳腺癌新辅助化疗疗效评估中的价值。方法采用免疫组化S-P法和荧光定量PCR方法检测32例"三阴"性乳腺癌患者新辅助化疗(CEF方案)前后乳腺癌组织BCSG1的表达,比较化疗前后肿瘤体积的变化情况,分析新辅助化疗前后BCSG1蛋白表达与肿瘤形态学变化的关系。结果23例乳腺癌患者新辅助化疗后肿瘤体积均有明显缩小,病灶缓解率(CR+PR)为84.4%;新辅助化疗后BCSG1 mRNA表达水平亦明显低于化疗前(P<0.05),BCSG1蛋白高表达率低于新辅助化疗前(P<0.01)。结论BCSG1分子和蛋白水平在"三阴"性乳腺癌新辅助化疗后均明显降低,与新辅助化疗后疗效呈负相关(r=-0.584,P<0.01),提示BCSG1可作为"三阴"乳腺癌新辅助化疗疗效的预测因子。

CK18对沉默HIF-1α的人“三阴”乳腺癌MDA-MB-231细胞凋亡的影响

目的研究CK18对HIF-1α干扰后的人"三阴"乳腺癌MDA-MB-231细胞凋亡的影响。方法采用shRNA干扰技术沉默MDA-MB-231细胞中的HIF-1α基因,选择RT-PCR及Western blot技术确定HIF-1α干扰效果。构建真核表达载体CK18-PEGFP-C1,以脂质体转染法将其转染入HIF-1α干扰后的MDA-MB-231细胞中,荧光显微镜下评估转染效率,Western blot检测CK18蛋白的表达情况。同时以空载体PEGFP-C1转染HIF-1α干扰后的MDA-MB-231细胞为阴性对照组。实验分别通过流式细胞术及Hoechst凋亡检测试剂盒检测CK18对细胞周期及凋亡的影响。结果构建了CK18-PEGFP-C1真核表达载体,基因测序结果与Gen Bank报道的完全一致,并转染了封闭HIF-1α基因的MDA-MB-231细胞,Western blot显示CK18蛋白表达上调。AnnexinⅤ-FITC/PI双染细胞凋亡检测分析CK18转染组早期凋亡率[(4. 9±0. 17)%]高于对照转染组[(3. 09±0. 25)%],差异有统计学意义(P <0. 05)。Hoechst染色试剂盒检测细胞凋亡显示CK18转染组的细胞凋亡率[(53±11)%]高于对照转染组[(21±13)%],差异有统计学意义(P <0. 05)。结论成功构建了CK18真核表达载体,并在HIF-1α干扰后的人"三阴"乳腺癌MDA-MB-231细胞中稳定表达后,可促进细胞凋亡。

A Prognostic Model Based on Colony Stimulating Factors-related Genes in Triple-negative Breast Cancer

Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells,playing an important role in the malignant progression of TNBC.This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes(CRGs),and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy.Methods We downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database.Through LASSO Cox regression analysis,we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score(CRRS).We further analyzed the correlation between CRRS and patient prognosis,clinical features,tumor microenvironment(TME)in both high-risk and low-risk groups,and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy.Results We identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model.Kaplan-Meier survival curves,time-dependent receiver operating characteristic curves,and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival,and the predictive ability of CRRS prognostic model was further validated using the GEO dataset.Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients.Moreover,patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil,ipatasertib,and paclitaxel.Conclusion We have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs,which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment.Moreover,the key genes within this model may represent potential molecular targets for future therapies of TNBC.

Effect of the Fu Zheng Method Applied by Hong-Feng CHEN on the Quality of Life in TNBC Patients and Empirical Research Based on Data Mining

Objective To preliminarily evaluate the efficacy of the Fu Zheng method(supporting resistance against pathogenic factors)applied by Professor Hong-Feng CHEN in improving the quality of life in patients with triple-negative breast cancer(TNBC)to collect and organize the traditional Chinese medicine(TCM)formulas applied by Professor Hong-Feng CHEN in the treatment of TNBC in order to explore the patterns in prescription.Methods The Functional Assessment of Cancer Therapy-Breast scale(FACT-B V4.0)were collected before and after treatment;the database and data mining platform for prescribed TCM formulas were constructed using Microsoft SQL Server 2005,and the patterns in drug pairing and combination were summarized by correlation analysis of data mining.Results The formulas improved the mean scores of the patients’physical well-being,social/family well-being,emotional well-being,physical function well-being and additional concerns(P<0.01);the drug combinations and pairs frequently used by Professor Hong-Feng CHEN were summarized.Conclusion The TCM formulas applied by Professor Hong-Feng CHEN can alleviate adverse physiological reactions,improve psychological conditions and improve function in patients.The formulas take spleen invigoration and stomach nourishment as well as blood circulation promotion and stagnation dissipation as the therapeutic principles,with simplicity in prescription and focus on care and protection the foundation of acquired constitution.

Augmented efficacy and the mechanism of a combined use of daunorubicin with rofecoxib in treatment of triple-negative breast cancer

Triple-negative breast cancer is the tumor that lacks expressions of estrogen receptor（ER）, progesterone receptor（PR） and human epidermal growth factor receptor-2（HER2）. A regular chemotherapy cannot eradicate triple-negative breast cancer. In the present study, we aimed to develop a combined use of daunorubicin and rofecoxib to treat triple-negative breast cancer, and reveal the underlying mechanisms. A gradient elution HPLC-UV method was developed for quantification, and the evaluations were performed on the triple-negative breast cancer MDA-MB-231 cells using a high content screening system. The results demonstrated that daunorubicin alone was insensitive to the triple negative breast cancer cells, while the combined use of daunorubicin and rofecoxib was able to effectively kill these triple-negative cancer cells, exhibiting a rofecoxib concentration-dependent manner. The mechanism revealed that the augmented anticancer efficacy was associated with direct killing effect, inducing apoptosis and inducing autophagy by the combination treatment. Besides, the apoptosis signaling pathways were correlated to a cascade of reactions by activating apoptotic enzyme caspase family and by suppressing anti-apoptotic gene expressed protein Bcl-2 family. In conclusion, this study provided a fundamental evidence for further developing the combined use of daunorubicin and rofecoxib formulation, hence offering a promising strategy for eradicating the triple negative breast cancer.

Roles of miRNA and lncRNA in triple-negative breast cancer

Triple-negative breast cancer(TNBC)is currently the most malignant subtype of breast cancer without effective targeted therapies,which makes its pathogenesis an important target for research.A growing number of studies have shown that non-coding RNA(ncRNA),including microRNA(miRNA)and long non-coding RNA(lncRNA),plays a significant role in tumorigenesis.This review summarizes the roles of miRNA and lncRNA in the progression,diagnosis,and neoadjuvant chemotherapy of TNBC.Aberrantly expressed miRNA and lncRNA are listed according to their roles.Further,it describes the multiple mechanisms that lncRNA shows for regulating gene expression in the nucleus and cytoplasm,and more importantly,describes lnc RNA-regulated TNBC progression through complete combining with miRNA at the post-transcriptional level.Focusing on miRNA and lncRNA associated with TNBC can provide new insights for early diagnosis and treatment—they can be targeted in the future as a novel anticancer target of TNBC.

Virtual screening for triple-negative breast cancer cell inhibitors based on telomere G-quadruplex structure

Triple-negative breast cancer is an aggressive subtype that frequently develops resistance to chemotherapy. It is expected to develop new anti-tumor drugs through targeting the structure of G-quadruplexes of the genes associated with this tumor. In this work, by targeting the 21-mer telomere G-quadruplex structure, compounds VB07 and VC02 were identified to stabilize the telomere G-quadruplex through structure-based high-throughput virtual screening. Cell cytotoxicity assay showed that VB07 and VC02 exhibited inhibitory effect on triple-negative breast cancer cells at the concentration of 5 μM. This study showed that structure-based high-throughput virtual screening was able to successfully identify the proper compounds targeting the telomere G-quadruplex, which exhibited inhibitory effects against the triple-negative breast cancer cells.