KLF5 suppresses TRAIL-induced cell apoptosis in PC-3 prostate cancer cells / 西安交通大学学报(医学版)

Objective: To explore the possibility of promoting tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis in prostate cancer PC-3 cell by inhibiting Krüppel-like factor 5 (KLF5). Methods: MTT assay, flow cytometry, Western blot assay and qRT-PCR assay were deployed to detect the cell viability, apoptosis and apoptotic markers in KLF5-inhibited and TRAIL-induced PC-3 cells. Results: After KLF5 was inhibited in TRAIL-induced PC-3 cells, cell viability reduced, apoptosis enhanced, the expressions of DR4 and DR5 increased while the expression of cellular fas-associated death domain-like interleukin-1β converting enzyme inhibitory protein (c-FLIP) decreased. Conclusion: Inhibiting KLF5 suppresses cell viability by promoting TRAIL-induced apoptosis in prostate cancer cell PC-3. It may be a potential means to treat hormone-insensitive prostate cancer.

Expression and clinical significance of TRAIL and c-FLIP in peripheral blood mononuclear cell of patients with rheumatoid arthritis / 中国免疫学杂志

Objective:To analyze the expression of TNF-related apoptosis-inducing factor TRAIL,c-FLIP and caspase-8 in peripheral blood mononuclear cells of patients with rheumatoid arthritis at different period,in order to provide a experimental basis that treating and looking for a more effective way and method.Methods:mRNA expression of TRAIL,c-FLIP and caspase-8 were detected by Real-time PCR from peripheral blood mononuclear cells;TRAIL,c-FLIP and caspase-8 were checked by Western blot from peripheral blood mononuclear cells.Results:The mRNA expression level of TRAIL,c-FLIP and caspase-8 from PBMC at different groups of RA:TRAIL mRNA in group M was 3.26±0.78 which was much higher than healthy controls(1.30±0.20) (P=0.028),and those of group L and group H (1.56±0.37 and 1.83±0.26 respectively) was slightly higher than controls(P<0.05).C-FLIP mRNA in low activity group (L),middle activity group (M) and high activity group (H) were 1.27±0.28,1.32±0.34 and 1.93±0.40 re-spectively,which was higher than that of healthy controls (1.08 ± 0.12) (P=0.035,0.034,0.030).The relative level caspase-8 mRNA in group L,group M,group H were(2.77 ±0.97),(4.52 ± 0.85),and(2.13 ± 0.44)which was higher than healthy controls (1.04±0.13) (P=0.023,0.012,0.032).The protein level of TRAIL,c-FLIP and caspase-8 in PBMC from different groups of RA patients:The expression of TRAIL in group M was significantly increased than group L,H and control(P<0.05) with no difference in group L.c-FLIP protein in all protein expressed in group M quantity highest,significantly higher than other groups.Caspase-8 expression in the group M and H was significantly enhanced than control (P=0.003,0.001 ) with no difference in group L (P> 0.05). Conclusion:During the different active stages of RA,in peripheral blood mononuclear cells,the expression of TRAIL,c-FLIP and caspase-8 are increased overall trend,possible can provide certain experimental reference for clinical therapy.

CRT promoted c-FLIP expression via NF-κB pathway in rheumatoid arthritis fibroblast-like synoviocytes / 天津医药

Objective To investigate the molecular mechanisms of upregulated expression of cellular Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein(c-FLIP)by calreticulin(CRT)in patients with rheumatoid arthritis (RA). Methods The semi-quantitative analysis and localization of c-FLIP in RA and osteoarthritis (OA)synovium were detected by immunohistochemistry.The fibroblast-like synoviocytes(FLS)were isolated by enzymatic digestion of synovial tissue specimens obtained from RA and OA patients,and cultured as an in vitro experiment model.The expressions of c-FLIP in RA and OA synovial fibroblasts were detected by immunofluorescence and Western blot assay. Whether CRT influenced c-FLIP expression and its molecular mechanism were explored by Western blot assay. Results The high expression of c-FLIP was found in RA synovium, mainly in the lining and sublining areas of FLS and vascular endothelial cells detected by immunohistochemistry.Meanwhile,weak staining of c-FLIP was observed in OA synovium.The expression of c-FLIP was significantly higher in RA synovium than that of OA synovium(t=11.717,P<0.001).Results of immunofluorescence and Western blot assay showed that c-FLIP was mainly located in cytoplasm, and which was higher expressed in FLS of RA than that of OA. The increased c-FLIP expression and phosphorylation of NF-κB were detected after being co-incubated with exogenous CRT (0, 10, 50, 100 μg/L), in dose-dependent manner. The effect of CRT upregulating c-FLIP expression was blocked by NF-κB inhibitor BAY 11-7082.Conclusion CRT can increase c-FLIP expression at least partly through NF-κB pathway in RA,which may provide therapeutic target for the treatment of RA.

Expression and significance of c-FLIP protein in sepsis mice with acute kidney injury / 中华危重病急救医学

Objective To observe the expression of cellular Fas-associated death domain-like interleukin-1β converting enzyme inhibit protein (c-FLIP) in sepsis mice with acute kidney injury (SAKI) and explore its significance. Methods Thirty male ICR mice were divided into the normal control group (Normal group), sham operation group (Sham group) and SAKI group by random number table method, with 10 mice in each group. The SAKI model of mice was established by cecal ligation and puncture (CLP); the Sham group was not ligated and the cecum was not punctured, and other surgical procedures were the same as the SAKI group; the Normal group did not experience any treatment. The serum and renal tissues of mice in each group were harvested 24 hours after CLP model establishment. The levels of serum creatinine (SCr) and blood urea nitrogen (BUN) were detected by enzyme linked immunosorbent assay (ELISA). The renal tissues were stained with hematoxylin-eosin (HE), and the pathological changes of renal tissues were observed under light microscope and the severity of injury was determined. The expression of c-FLIP in renal tissues was detected by immunohistochemistry. The expression of c-FLIP, Bax and caspase-3 protein in renal tissue was detected by Western Blot. The correlation between c-FLIP expression and Bax, caspase-3 protein expressions in renal tissues were analyzed by Pearson test. Results In the Normal group and the Sham group, the renal tubular epithelial cells were regular and intact, and no interstitial inflammatory cell infiltration was observed; the renal injury score was both 1.30±0.48; immunohistochemistry showed a large amount of c-FLIP positive expression in renal tubular epithelial cells (IA: 120.20±3.87, 116.70±3.46); Western Blot showed high expression of c-FLIP in renal tissues (c-FLIP/GAPDH: 0.99±0.01, 0.98±0.02), and low expressions of Bax and caspase-3 (Bax/GAPDH: 0.16±0.04, 0.19±0.03, caspase-3/GAPDH: 0.24±0.04, 0.23±0.05). Compared with the Sham group, in the SAKI group, renal tubular epithelial cells were degenerated and necrosis, and a large number of interstitial inflammatory cells infiltrated, the renal injury score was significantly increased (4.60±0.52 vs. 1.30±0.48, P < 0.01); the levels of SCr and BUN were significantly increased [SCr (μmol/L): 193.90±13.54 vs. 24.50±3.78, BUN (mmol/L): 81.60±7.26 vs. 5.20±0.92, both P < 0.01]; the c-FLIP positive cells in renal tissues was significantly reduced (IA: 17.11±0.82 vs. 116.70±3.46, P < 0.01); the expression of c-FLIP protein in renal tissues was significantly decreased (c-FLIP/GAPDH: 0.29±0.03 vs. 0.98±0.02, P < 0.01), while the expressions of Bax and caspase-3 protein were significantly increased (Bax/GAPDH: 0.87±0.06 vs. 0.19±0.03, caspase-3/GAPDH: 0.88±0.07 vs. 0.23±0.05, both P < 0.01]. The correlation analysis showed that the c-FLIP protein was significantly negatively correlated with Bax (r = -0.468, P = 0.029) and caspase-3 protein expressions (r = -0.663, P = 0.004). Conclusions The expression level of c-FLIP protein was significantly down-regulated in renal tissue of SAKI, and its down-regulation mechanism was associated with increased apoptosis of renal tubular epithelial cells, which could be an effective target for the treatment of SAKI.

L-carnitine sensitizes human glioblastoma cells to TRAIL-induced apoptosis / 中国药理学通报

Aim To investigate the enhancing effect of L-carnitine as a sensitizer on tumor necrosis factor-re-lated apoptosis inducing ligand ( TRAIL)-induced ap-optosis in glioma cells. Methods Glioma cell U87 was used as model cell line. Cell viability was determined by CCK-8 , and apoptosis was assessed by Annexin V-FITC/PI staining, caspase-3 activity and expres-sion. The expression and transcription of nuclear factor kappa B ( NF-κB ) and FLICE inhibiting protein ( c-FLIP) were measured by RT-PCR and Western blot. In addition, NF-κB was knockdown to analyze its regu-lating effect on c-FLIP expression. Results The com-bination treatment with TRAIL and L-carnitine signifi-cantly inhibited cell proliferation and induced apopto-sis. Compared with control, combinational treatment significantly suppressed the transcription and expres-sion of c-FLIP as well as translocation of NF-κB. Through silencing NF-κB, NF-κB was found to act as upstream signaling to regulate c-FLIP. Conclusion L-carnitine sensitizes TRAIL-induced tumor cell apoptosis via suppression of NF-κB-dependent c-FLIP expres-sion.

The Histone Deacetylase Inhibitor Trichostatin A Sensitizes Human Renal Carcinoma Cells to TRAIL-Induced Apoptosis through Down-Regulation of c-FLIP(L)

Histone acetylation plays a critical role in the regulation of transcription by altering the structure of chromatin, and it may influence the resistance of some tumor cells to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) by regulating the gene expression of components of the TRAIL signaling pathway. In this study, we investigated the effects and molecular mechanisms of trichostatin A (TSA), a histone deacetylase inhibitor, in sensitizing TRAIL-induced apoptosis in Caki human renal carcinoma cells. Our results indicate that nontoxic concentrations of TSA substantially enhance TRAIL-induced apoptosis compared with treatment with either agent alone. Cotreatment with TSA and TRAIL effectively induced cleavage of Bid and loss of mitochondrial membrane potential (MMP), which was associated with the activation of caspases (-3, -8, and -9) and degradation of poly (ADP-ribose) polymerase (PARP), contributing toward the sensitization to TRAIL. Combined treatment with TSA and TRAIL significantly reduced the levels of the cellular Fas-associated death domain (FADD)-like interleukin-1beta-converting enzyme (FLICE) inhibitory protein (c-FLIP), whereas those of death receptor (DR) 4, DR5, and FADD remained unchanged. The synergistic effect of TAS and TRAIL was perfectly attenuated in c-FLIP(L)-overexpressing Caki cells. Taken together, the present study demonstrates that down-regulation of c-FLIP contributes to TSA-facilitated TRAIL-induced apoptosis, amplifying the death receptor, as well as mitochondria-mediated apoptotic signaling pathways.

Relationship between c-FLIP protein and drug resistance of ovarian cancer cells to TRAIL-induced apoptosis / 西安交通大学学报(医学版)

Objective To explore the mechanism of drug resistance of ovarian cancer cells to TRAIL-induced apoptosis.Methods We collected 3AO cells and CAOV3 cells,respectively,at 18,24,48 and 72 hour under 12.5,25,50 and 100 ng/mL concentrations of TRAIL.The rate of cell growth inhibition was checked by methyl thiazolyl tetrazolium (MTT)assay to evaluate the effect of TRAIL.Morphology of apoptotic cells was observed by TdT-mediated-dUTP nick end labeling (TUNEL).The apoptosis rate was detected by flow cytometry (FCM)and C-FLIP protein was determined by Western blotting.Results TRAIL inhibited the growth of 3AO and CAOV3 cells.The rate of growth inhibition at 24 hour was 28% in 3AO cells and 10% in CAOV3 cells.TRAIL induced apoptosis of cells.The apoptosis rate at 24 hour was 8.5% in 3AO cells,which was higher than 5.5% in CAOV3 cells.The expression level of C-FLIP protein was higher in CAOV3 cells than in 3AO cells.Conclusion C-FLIP protein is an important protein that regulates drug resistance of ovarian cancer cells to TRAIL-induced apoptosis.

Studies on cell death and survival regulatory protein c-FLIP as a target for cancer treatment / 中国药理学通报

Many tumor cells are resistant to cell apoptosis through the expression of antiapoptotic proteins. c-FLIP is a ma-jor resistance protein of antiapoptosis. In human cells, there are three types of c-FLIP, c-FLIPL , c-FLIPS and c-FLIPR . The c-FLIP binds to FADD to prevent the formation of procaspase-8-DISC and the subsequent activation of caspase cascade. Further-more, c-FLIPL and c-FLIPS have multifunctional roles in various cellular signaling pathways, as well as up-regulating several cyto-protective signaling. Studies show that upregulation of c-FLIP has been found in various tumors, and its downregulation has been shown to restore apoptosis triggered by various chemothera-peutic agents, like the transcriptional regulating agents, trichos-tatin-A, camptothecin, cisplatin, doxorubicin, etc. or other new biotechnologies, such as the specific siRNA. Therefore, c-FLIPS are important targets of cancer therapy. This review summarizes the results on the role of c-FLIP in cancer chemotherapy of tradi-tional antitumor agents and siRNA, and to provide new ideas and rationales of searching for the antiapoptosis effective compounds that can specifically antagonize c-FLIP.

Kaurene diterpene induces apoptosis in U87 human malignant glioblastoma cells by suppression of anti-apoptotic signals and activation of cysteine proteases

Gliomas are the most common and malignant primary brain tumors in humans. Studies have shown that classes of kaurene diterpene have anti-tumor activity related to their ability to induce apoptosis. We investigated the response of the human glioblastoma cell line U87 to treatment with ent-kaur-16-en-19-oic acid (kaurenoic acid, KA). We analyzed cell survival and the induction of apoptosis using flow cytometry and annexin V staining. Additionally, the expression of anti-apoptotic (c-FLIP and miR-21) and apoptotic (Fas, caspase-3 and caspase-8) genes was analyzed by relative quantification (real-time PCR) of mRNA levels in U87 cells that were either untreated or treated with KA (30, 50, or 70 µM) for 24, 48, and 72 h. U87 cells treated with KA demonstrated reduced viability, and an increase in annexin V- and annexin V/PI-positive cells was observed. The percentage of apoptotic cells was 9% for control cells, 26% for cells submitted to 48 h of treatment with 50 µM KA, and 31% for cells submitted to 48 h of treatment with 70 µM KA. Similarly, in U87 cells treated with KA for 48 h, we observed an increase in the expression of apoptotic genes (caspase-8, -3) and a decrease in the expression of anti-apoptotic genes (miR-21 and c-FLIP). KA possesses several interesting properties and induces apoptosis through a unique mechanism. Further experiments will be necessary to determine if KA may be used as a lead compound for the development of new chemotherapeutic drugs for the treatment of primary brain tumors.

Research progress on the dual regulation of c-FLIP in apoptosis and proliferation and the relationship between c-FLIP and tumor prognosis, chemotherapy, and TRAIL treatment in cancers / 中国肿瘤临床

Cellular Fas-associated death domain-like interleukin-1β-converting enzyme inhibitory protein (c-FLIP) belongs to the death effector domain superfamily, which is important in regulating apoptosis and proliferation. c-FLIP inhibits the extrinsic recep-tor-mediated apoptotic pathways and intrinsic mitochondrial apoptotic pathways through competition with caspase-8 for recruitment to Fas-associated death domain protein. Moreover, the cleavage products (i.e., p43-FLIP fragment and p22-FLIP fragment) directly acti-vate NF-κΒ, Erk survival signaling, and other non-apoptotic signaling pathways. The c-FLIP (L) can function either as an anti-apoptotic molecule, in a way analogous to c-FLIP (S) and c-FLIP (R), or as a pro-apoptotic molecule to facilitate the activation of caspase-8 at the death-induced signaling complex. The identified dual functionality of c-FLIP depends on various factors, including its expression level, interaction with caspase-8, and its subcellular localization. c-FLIP is frequently over-expressed in many different tumor types, and con-tributes to tumor cell immune surveillance, chemotherapy resistance, and apoptosis-resistance induced by TNFα, TRAIL, and FasL. Fur-thermore, c-FLIP is essential in obtaining aggressive biological behaviors, and is useful in predicting the prognosis of patients with vari-ous malignant tumors. This review focuses on the molecular mechanisms that control the dual regulation of c-FLIP in life/death deci-sion at death-induced signaling complex. Increasing evidence supports the function of c-FLIP as a tumor therapeutic marker to restore an apoptotic response for TRAIL therapy in cancers. Insight into these processes will improve our understanding of apoptosis, and pro-vide new approaches for rational treatment strategies.

MiR-512-3p expression pattern and function in breast cancer / 中国肿瘤临床

Objective:This study aims to investigate the expression pattern of miR-512-3p in breast cancer and noncancerous paired specimens as well as the effects of miR-512-3p on the proliferation, apoptosis, cell cycle, and cloning of MD-MBA-231 breast cancer cells. The study also aims to identify the miR-512-3p target gene. Methods:Reverse transcription-polymerase chain reaction (RT-PCR) was conducted to quantify the miR-512-3p expression in breast cancer and noncancerous paired specimens. Methylthiazol tetrazolium (MTT) assay, flow cytometry, and clone formation assay were used to characterize the function of miR-512-3p in breast cancer. Target prediction was performed using the TargetScan, PicTar, and miRanda software. The results were validated using RT-PCR and western blot target validation. Results:The relative expression of miR-512-3p in breast cancer specimens was significantly lower than those in normal breast specimens (P<0.05). MTT assay revealed that 48 h after transfection, miR-512-3p significantly repressed the proliferation of MD-MBA-231 cells at a suppression rate of 45.38%and at a concentration of 100 nmol/L. MiR-512-3p increased the percentage of early apoptotic cells in the treatment groups (9.32 ± 0.41)%compared with those in the blank controls (3.1 ± 0.54)%and in the negative controls (2.9 ± 0.39)%(P<0.05). Significant differences were found in the percentages of the G0/G1-and G2/M-phase cells after miR-512-3p transfection compared with those in the controls (P<0.05). In the cloning assay, clone formation was inhibited in the miR-512-3p-transfected groups compared with those in the control groups. RT-PCR and western blot results indicate that miR-512-3p significantly inhibited the c-FLIP mRNA and protein expression. Conclusion:MiR-512-3p expression is relatively decreased in breast cancer specimens compared with those in the normal samples. The negative effect of miR-512-3p on cell proliferation and clone formation and its positive effect on early apoptosis through c-FLIP targeting suggest that miR-512-3p acts as a tumor suppressor gene in breast cancer. Therefore, miR-512-3p may be a new target for the diagnosis and treatment of breast cancer in the future.

c-FLIP antisense oligonucleotide-loaded nanoparticles inhibit growth of human orbitalrhabdomyosarcoma ienograft in nude mice / 第二军医大学学报

Objective To investigate the effect of cellular Fas-associated death domain-like interleukin-1β-converting enzyme-inhibitory protein (c-FLIP) antisense oligonucleotide (ASODN)-loaded nanoparticles (NP) on the human orbital rhabdomyosarcoma xenograft in nude mice, so as to assess the feasibility of nanoparticles as a gene vector. Methods The model of human orbital rhabdomyosarcoma xenograft was established in nude mice, and the tumors were injected with c-FLIP ASODN NP, c-FLIP ASODN or normal saline (NS). The tumor volume and histopathological changes of tumor were observed. Western blotting analysis and immunohistochemical analysis were used to examine the expression of c-FLIP in tumor tissues of each group. Apoptosis of tumor cells was detected using TUNEL method. Results The growth of human orbital rhabdomyosarcoma in nude mice was significantly inhibited in ASODN NP group compared with the other two groups. Western blotting analysis showed that c-FLIP protein expression in ASODN NP and ASODN groups was significantly decreased compared with NS group (P<0. 05). Immunohistochemical study showed that c-FLIP expressionwas found in the endochylema, and the c-FLIP positive cells in ASODN NP group was significantly less than those in the other two groups (P<0.05). Tumor cell apoptosis was observed in both ASODN NP and ASODN groups, with more found in the former, and only a few apoptotic cells were found in the NS group. Conclusion c-FLIP ASODN NP can effectively inhibit the growth of human orbital rhabdomyosarcoma xenograft in nude mice, indicating that nanoparticles may serve as a safe and effective vector for ASODN.

Quinazolinone exposure induces apoptosis and changes expressions of Fas/FasL and C-Flip in embryonic mice testicles.

Quinazolinones represent a class of sedative and anticancer drugs. Quinazolinones-based compounds have ability to suppress prostate tumor growth via apoptosis. Apoptosis is very common in embryos and adults of normal and injured mammalian testes. Effects a new derivative of quinazolinone (4(3H) quinazolinone-2-ethyl-2-phenyl ethyl (QEPE)), on the testis of Balb/C mice embryos were investigated. QEPE was able to reduce number of germ cells and diameter of seminiferous tubules. TUNEL assay analysis indicated that reduction correlated with an increase in the number of apoptotic cell. Furthermore, electron microscope observations confirmed typical apoptotic morphologies characterized by chromatin fragmentation. Finally, RT–PCR analysis showed QEPE increases the levels of Fas/Fasl and decreases C-Flip mRNAs in the testis of exposed embryos.

Expression of c-FLIP in Gastric Cancer and its Relation to Tumor Cell Proliferation and Apoptosis

BACKGROUND: The expression of c-FLIP (cellular Fas-associated death domain-like interleukin-1 beta-converting enzyme (FLICE)-inhibitory protein), which is a member of the family of inhibitors of apoptosis, has been associated with tumor development and progression. The aim of this study was to evaluate the expression of c-FLIP in gastric cancer and its correlation with tumor cell proliferation, apoptosis and the clinicopathologic features. METHDOS: Immunohistochemical staining with anti-c-FLIP antibody was performed in 98 tissue samples obtained from gastric cancer patients who underwent surgical treatment. The apoptotic cells were visualized by terminal deoxynucleotidyl transferase (TdT) mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL), and the proliferative cells were visualized by staining with Ki-67 antibody. RESULTS: The positive expression of c-FLIP in the gastric cancer tissues was demonstrated in 57.1% of the cases. The expression of c-FLIP was increased in the gastric cancer tissues compared with the matched normal gastric mucosa. The expression of c-FLIP was significantly associated with histologic differentiation (p=0.038). However, there was no association between the c-FLIP expression and the other clinicopathological parameters, including patient survival. The Ki-67 labeling index (KI) for the 98 tumors ranged from 7.6 to 85.0 with a mean KI of 50.4+/-15.7. The mean KI value of the c-FLIP positive tumors was 54.1+/-15.3 and this was significantly higher than that of the c-FLIP negative tumors (p=0.005). The apoptotic index (AI) for the 98 tumors ranged from 0.0 to 10.0 with a mean AI of 7.4+/-2.3. There was no significant difference between the c-FLIP expression and the AI (p=0.347). CONCLUSIONS: These results suggest that the c-FLIP expression may be associated with tumor cell proliferation of gastric cancer.

c-FLIP:The Regulator of Extrinsic Apoptotic Pathway / 生物化学与生物物理进展

Cellular FADD-like interleukin-1-? converting enzyme inhibitory protein (c-FLIP) is a kind of inhibitory protein of caspase with the death effector domain (DED), naturally existing in many species such as virus, eukaryote and mammal inclusively. Recently, it has been discovered that c-FLIP participates the regulation of apoptosis. Overexpression of c-FLIP may inhibits the apoptosis induced by the death receptor of Fas and TRAIL-R. With the development on the mechanism of action and molecular regulation of c-FLIP, its multi-biology function has been found, and also it is associated with the nosogenesis and progression of many diseases.

Study of the therapeutic time window and dose response effects of epirubicin on the expression of c-FLIP in breast cancer / 中国普通外科杂志

Objective To investigate the therapeutic time window and the dose response effects of epirubicin on the expression of c-FLIP in breast cancer.Methods MCF-7and MDA-MB-231 breast cancer cells were divided into two groups: epirubicin groups were treated with 4.0,2.0,1.0,0.5 and 0.25mg/L of epirubicin,and control groups were treated with 0.9% sodium chloride solution at the same dose.After treatment for 24,48 and 72 h,the incubated cells were collected for the measurement of c-FLIP by RT-PCR,and for examination of percent of apoptosis cells with flow cytometry.ResultsA dose-time-dependent pattern was observed.The expression of c-FLIP in MCF-7and MDA-MB-231 breast cancer cell lines declined gradually as the epirubicin concentration increased and treatment time was prolonged.Percentage of apoptosis breast cancer cells increased gradually as the epirubicin concentration was increased and treatment time was prolonged,and percentage of apoptosis cells was the highest when breast cancer cells were treated with 2 mg/L epirubicin for 72 h.ConclusionsEpirubicin can promote apoptosis of breast cancer cells by inhibiting the expression of c-FLIP,and its inhibitory effect is most pronounced when breast cancer cells are treated with 2 mg/L epirubicin for 72 h.