Objective To explore effect of DLL4 gene in MCF-7 cells of human breast cancer which was inhibitted by short hair in RNA （shRNA） on inducing apoptosis and chemosensitivity to docetaxel. Methods Specific shRNA was designed in accordance with DLL4 gene and transfected into MCF-7 cells of human breast cancer with liposomal （Lip-shRNA group）， MCF-7 cells transfected with only liposomal as Lip group， and control group without any treatment. Expressions of DLL4 protein in 3 groups were detected by immunohistochemical method， apoptosis and cell cycle distribution were examined by flow cytometry （FCM）. Proliferations and sensitivity of MCF-7 cells to docetaxel in 3 groups were determined by methylthiazoyl-tetrazolium bromide （MTT）. Results The averages optical density value and rate of positive area of Lip-shRNA group were significantly lower than that of other 2 groups （P＜0.01）. The levels of A value at 24h， 48h， and 72h in Lip-shRNA group were significantly lower than that of other 2 groups （P＜0.01）. Rates of cell apoptosis at 24h， 48h， 72h and 96 h in Lip-shRNA group were significantly higher than that of other 2 goups （P＜0.05），and ratio of G2/M was higher too （P＜0.05）. IC50 of Lip-shRNA group to docetaxel was significantly lower than that of other 2 groups （P＜0.05）. Conclusions The RNA interference technology can effectively block the expression of DLL4 gene. Inhibition of DLL4/Notch signaling pathway can lead to proliferation inhibition of cancer cell and a concomitant increase in cells undergoing apoptosis， and can enhance the cell sensitivity to docetaxel. DLL4 may be an important target for therapeutic approach of breast cancer.
Objective To study the expressions of Delta-like ligand 4 （DLL4） and S100A4 in breast carcinoma of differect molecular subtypes and explore its clinical significance. Methods The expressions of DLL4 and S100A4 in all molecular subtypes were tested by SP immunohistochemistry in 108 cases of breast carcinoma and 40 cases of paracancerous tissues from Taihe Hospital. The Luminal A， Lumianl B， HER-2 over-expression， and basal-like subtypes was 51， 26， 17， and 14 cases， respectively. Then the correlation of DLL4 and S100A4 expression with patients’ clinical and pathological features were analyzed. Results The positive expression rates of DLL4 and S100A4 in breast carcinoma was 67.6%（73/108）and 62.0%（67/108）respectively， which were significantly higher than those in paracancerous tissues〔22.5%（9/40） and 45.0%（18/40）〕， P＜0.05. The positive expression rates of DLL4 and S100A4 in breast carcinoma tissues of HER-2 over-expression and basal-like subtypes were significantly higher than those in breast carcinoma tissues of Luminal A and Luminal B subtypes （P＜0.05）. The expressions of DLL4 and S100A4 in breast carcinoma tissues with lymph node metastasis and without lymph node metastasis were significantly different（P＜0.05）. There was positiver elationship between the expressions of DLL4 and S100A4 in breast carcinoma tissues（rs=0.217，P＜0.01）. Conclusions DLL4 and S100A4 are highly expressed in breast carcinoma tissues of HER-2 over-expression and basal-like subtypes， and are all related with prognosis of breast carcinoma. These results suggest that they might be important factors in breast carcinogenesis and tumor development， metastasis. These proteins are indicators of metastasis and predictors for prognosis of breast carcinoma.
ObjectiveTo test the expressions of human mammary gland globin (hMAM) mRNA in the peripheral blood of breast cancer and breast benign lesions patients, try to provide the theory basis for the choice of breast cancer molecular marker. MethodsPolymerase chain reaction (PCR) technology was used to detect the expressions of hMAM mRNA in peripheral blood of 78 cases of breast cancer patients, 15 cases of hyperplasia of mammary gland, and 15 cases of breast fibroadenoma. The relationship between the expressions of hMAM-mRNA in peripheral blood of breast cancer patients with patient's age, tumor size, pathological type, tumor stage, axillary lymph node metastasis, and the ER, PR and HER-2 status were analyzed. ResultsThe expressions of hMAM-mRNA in peripheral blood were not detected in breast hyperplasia and breast fibroadenoma patients, but the peripheral blood hMAM-mRNA expression rate in breast cancer patients was 48.72% (38/78), the difference was statistically significant (χ2=12.357, P=0.000). The expression of peripheral blood hMAM mRNA was not related to the patient's age, tumor size, pathological type, and ER, PR and HER-2 status (P > 0.05), but the expression of peripheral blood hMAM mRNA was related to the clinical staging of tumor (Z=-2.214, P=0.027) and lymph node metastasis status (Z=-2.754, P=0.006). ConclusionPeripheral blood hMAM-mRNA detected is a sign of breast cancer, further research is needed to confirm whether hMAM mRNA detection in peripheral blood correlates with poor prognosis of breast cancer patients.
ObjectiveTo explore expressions of the metaherin (MTDH) mRNA and its protein in hepatitis B related hepatocellular carcinoma tissues and its clinical significance. MethodsSeventy two tissues of patients with hepa-titis B related hepatocellular carcinoma who were treated in Affiliated Taihe Hospital of Hubei Medical University from Jul. 2012 to Oct. 2013 were collected retrospectively. Quantitative PCR (Q-PCR) and Western blot methods were used to detect the expression levels of MTDH mRNA and its protein in 10 cases of hepatitis B related hepatocellular carcinoma tissues and pericarcinoma tissues. Besides, immunohistochemistry was used to determine the expression of MTDH protein in 72 cases of hepatocellular carcinoma tissues, then the relationship of expression of MTDH protein and clinico-pathological features was explored. ResultsThe expression levels of MTDH mRNA and its protein in hepatocellular carcinoma tissues were both higher than those of pericarcinoma tissues (8.50±0.84 vs. 4.55±0.81, t=10.797, P=0.000; 0.65± 0.24 vs. 0.25±0.16,t=6.375, P=0.013). The MTDH protein was positively expressed in 42 cases (58.3%) and negatively expressed in 30 cases (41.7%) of hepatocellular carcinoma tissues. The results of logistic regression showed that,MTHD protein was up-regulated in patients with category Ⅲ of Edmondson grade (OR=4.783, 95% CI:2.663-11.918, P=0.020), microvascular invasion (OR=37.790, 95% CI:2.227-99.434, P=0.005), and lymph node metastasis (OR=7.332, 95% CI:3.325-30.669, P=0.023). ConclusionExpressions of MTDH mRNA and its protein are both higher in hepatitis B related hepatocellular carcinoma tissue, which are correlated with poor prognosis.
ObjectiveTo explore expression of Mdm2 in the estrogen receptor α (ERα)-positive breast cancer tissues and fibroadenoma of breast tissues, and to explore the effect of MDM2-siRNA on cell proliferation, colony formation, and apoptosis for MCF-7 cells. Methods① Seventy eight ERα-positive breast cancer patients identified by histopathological examination, who underwent surgery in our hospital from June 2012 to October 2015, as well as 10 fibroadenoma of breast patients underwent surgery in the same period, were collected retrospectively to determine the expression of Mdm2, then explore the relationship between the expression of Mdm2 and clinical pathological characteristics of ERα-positive breast cancer patients. ② MCF-7 cells were divided to MDM2-siRNA group (added with MDM2-siRNA), negative control group (added with negative siRNA), and blank control group (added without any reagent). Expression of Mdm2, cell proliferation rate, number of colony formation, and apoptosis rate were determined in the MCF-7 cells of 3 groups. Results① No one of fibroadenoma of breast patients was found positive expression of Mdm2 (0/10), and 38 of 78 ERα-positive breast cancer patients were found the positive expression of Mdm2 (48.7%), which is higher than that of fibroadenoma of breast tissues (χ2=12.357, P=0.000). In ERα-positive breast cancer patients, expression of Mdm2 was related with TNM staging and number of metastasic lymph node (P < 0.050), the positive expression rate of Mdm2 was higher in patients with later TNM staging or more metastasic lymph node. ② Cell proliferation rates on 2, 3, and 4 days after transfection, expression level of Mdm2, and number of colony formation were all lower (P < 0.050), but the apoptosis rate was higher in MDM2-siRNA group (P < 0.050), comparing with negative control group and blank control group. But there was no significant difference between negative control group and blank control group on aforementioned indexes (P > 0.050). ConclusionMdm2 is a diagnostic marker in ERα-positive breast cancer patients, and treatment targeting it might has a certain therapeutic value.
Objective To systematically review the correlation between polymorphism of DNA methyltransferase 1(DNMT1) rs16999593 and the susceptibility of breast cancer. Methods Databases such as PubMed, EMbase, Web of Science, Chinese Biomedical Literature Database, CNKI, WanFang, and VIP database were searched from inception to Mar. 2017 to collect case-control studies on the correlation between DNMT1 rs16999593 C/T polymorphism and the susceptibility of breast cancer. Two reviewers independently identified the literatures according to inclusion and exclusion criterias, extracted data, and assessed the quality of the included studies. The meta-analysis was performed by using RevMan 5.3 software. Results A total of 5 studies involving 1 741 cases and 1 917 control subjects were included. The results of meta-analysis showed that, dominate model ［TT+TC vs. CC: OR=0.63, 95% CI was (0.30, 1.30), P=0.21］, homozygous model ［TT vs. CC: OR=1.01, 95% CI was (0.70, 1.47), P=0.95］, heterozygous model ［TC vs. CC: OR=0.44, 95% CI was (0.18, 1.04), P=0.06］, and additive model ［T vs. C: OR=1.29, 95% CI was (0.90, 1.86), P=0.16］ were not significantly related to breast cancer, but recessive gene model was related to breast cancer ［TT vs. TC+CC: OR=1.74, 95% CI was (1.01, 3.00), P=0.04］. Conclusion The current studies showed that, DNMT1 rs16999593 TT genotype decreases the susceptibility of breast cancer.
ObjectiveTo study the infiltration situation of breast cancer surface skin, and explore the characteristics of infiltration of breast cancer surface skin at the molecular level. MethodsNested reverse transcription-polymerase chain reaction technique was used to detect the expressions of human mammaglobin(hMAM)mRNA in 15 cases of hyperplasia of mammary gland tissues, 15 cases of breast fibroadenoma tissues, and 60 cases of breast cancer tissues and their corresponding tumor surface skins. The relationship of the hMAM mRNA expression in the surface skins of breast cancer tissue to its clinicopathologic characteristics was analyzed. ResultsThe hMAM mRNA positive expressions in the breast fibroadenoma tissues, hyperplasia of mammary gland tissues, and breast cancer tissues were 40.00%(6/15), 53.33%(8/15), and 83.33%(50/60), respectively, which in the breast cancer tissues was significantly higher than that in the breast fibroadenoma tissues or hyperplasia of mammary gland tissues(P < 0.05). There was no hMAM mRNA positive expression in the surface skin of fibroadenoma or hyperplasia of mammary gland tissues, but there was 3(5.00%)cases of the hMAM mRNA positive expressions in the breast cancer surface skin. The hMAM mRNA positive expression in the breast cancer surface skin was not related with the patient age, tumor diameter, and tumor staging(P > 0.05), but was related with axillary lymph nodes metastasis and distance from tumor to nipple less than 4 cm(P < 0.05). ConclusionsThe hMAM mRNA highly expresses in breast cancer tissue and it has a certain value in the diagnosis of infiltration of breast cancer surface skin. The patients with axillary lymph node metastasis and distance from tumor to nipple less than 4 cm are more susceptible to infiltration of breast cancer surface skin.
Objective To investigate the expression of hypoxia inducing factor 1 alpha (HIF-1α) in human breast cancer and its relationships with microvessel density （MVD）, proliferating cell nuclear antigen (PCNA) protein, other tumor biomarkers and clinicopathologic factors. Methods Immunohistochemical staining (SP) was used to measure the expressions of HIF-1α and PCNA in human breast fibroadenoma, usual hyperplasia and adenocarcinoma, and the MVD was determined by anti-CD34 immunostaining. Results No HIF-1α was observed in the lesions of breast fibroadenoma and hyperplasia. However, the positive expression rate of HIF-1α in the ductal carcinoma in situ （DCIS） was 55.0% (11/20) and the infiltrative breast cancer was 85.0%(51/60). The total high expression rate of PCNA in breast cancer was 75.0% (60/80), in which the rate of DCIS counted for 65.0% (13/20) and the rate of infiltrative adenocarcinoma counted for 78.3% (47/60). There were positive correlations between the expresson of HIF-1α and the expression of PCNA (r=0.693， P＜0.01) and MVD in DCIS (r=0.682, P＜0.05), respectively, but there was no relation between HIF-1α and MVD in infiltrative breast cancer. The expression of HIF-1α was associated with tumor cell proliferation, lymph node metastasis, estrogen receptor status (P＜0.01). Conclusion The expression of HIF-1α increased in breast cancer and it is associated with tumor cell proliferation, lymph node metastasis, estrogen receptor status. Thus, HIF-1α may play an important role in the tumor cell proliferation, vasiformation, progression and metastasis of breast cancer, and may become a new target for tumor treatment.