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In recent years, the incidence of extrahepatic cholangiocarcinoma (ECC) has been increasing annually. As a result of frequently invading adjacent structures, such as hepatic artery, hepatic vein, and portal vein, and low radical resection rate, the prognosis is poor. Even if radical resection is completed early, the 5-year survival rate is still less than 30%. At present, whether postoperative adjuvant therapy can improve the prognosis of ECC remains a research hotspot and a controversial point. This article will combine the latest research results to discuss the plan and status of postoperative adjuvant therapy after ECC, as well as analyze the effect of postoperative adjuvant therapy on ECC.
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@#Objective To evaluate the efficacy and safety of Huanglian Wendan Decoction (黄连温胆汤, HLWDD) alone or combined with western medicine in treating insomnia caused by phlegm-heat internal disturbance in recent 10 years. Methods The randomized controlled trials of HLWDD alone or combined with western medicine in treating insomnia caused by phlegm-heat internal disturbance from January 1, 2012 to April 1, 2022 were searched in China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), Wanfang Database, China BioMedical Literature Database (CBM), PubMed, Web of Science, Embase, and Cochrane Library databases. After being screening, the included literature was analyzed to evaluate the effective rate, Pittsburgh Sleep Quality Index (PSQI) score, traditional Chinese medicine (TCM) syndrome score, and adverse reactions of HLWDD on insomnia caused by phlegm-heat internal disturbance. The subgroup analyzed the effect of HLWDD after different treatment courses, and compared the therapeutic effects of HLWDD alone and HLWDD combined with western medicine. Results Twenty-seven randomized controlled trials were finally included, with a total of 2 395 patients. The results of the meta-analysis showed that the curative effect of HLWDD alone or combined with the western medicine group was better than that of the western medicine group [RR = 1.14, 95% CI (1.06, 1.22), P = 0.000]. The PSQI score [SMD = – 0.31, 95% CI (– 0.42, – 0.20), P = 0.000], TCM syndrome score [SMD = – 0.40, 95% CI (– 0.67, – 0.12), P = 0.005], and adverse reaction rate [RR = 0.21, 95% CI (0.15, 0.29), P = 0.000] of HLWDD alone or combined with western medicine group were significantly reduced compared with the western medicine group. The subgroup’s analysis showed that the curative effect of HLWDD alone or combined with western medicine group of 4 weeks treatment course was better than that of the western medicine group [RR = 1.14, 95% CI (1.03, 1.26), P < 0.05]. The TCM syndrome score of HLWDD alone or combined with the western medicine group of 4 weeks treatment course decreased more obviously than that of the western medicine group [SMD = – 0.60, 95% CI (– 0.96, – 0.25), P < 0.05]. There were no significant differences between HLWDD alone or combined with western medicine group and western medicine group with different treatment courses based on PSQI score and adverse reaction rate. Based on the effective rate, the comparison between the HLWDD alone group and the western medicine group [RR = 1.09, 95% CI (1.00, – 1.20) P < 0.05], and between the HLWDD combined with western medicine group and the western medicine group [RR = 1.15, 95% CI (1.03, 1.29), P < 0.05] was the same. PSQI score [SMD = – 0.44, 95% CI (– 0.59, – 0.30), P < 0.05] and TCM syndrome score [SMD = – 1.10, 95% CI (– 1.59, – 0.61), P < 0.05] of HLWDD combined with western medicine group were significantly lower than those of the western medicine group. There were no significant differences of adverse reaction rate between HLWDD alone group [RR = 0.08, 95% CI (0.04, 0.17), P < 0.05] and HLWDD combined with western medicine group [RR = 0.36, 95% CI (0.24, 0.53), P < 0.05]. Conclusion HLWDD alone or combined with western medicine is an effective treatment for insomnia caused by phlegm-heat internal disturbance, which has a high effective rate, significantly reduced PSQI score and TCM syndrome score, and favorable safety. The best course of treatment is 4 weeks.
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Objective To explore the value of an innovative teaching model which combined problem based learning (PBL) method with case based learning (CBL) method in clinical oncology Teaching.Methods 68 students were divided into the combinational teaching group (30 cases) and the LBL group (38 cases).The combinational teaching group was taught by PBL method combined with CBL method, and this dual track teaching was based on cases and problems.The traditional teaching group was taught by LBL method.The teaching effect was evaluated by students' questionnaire survey and test score.SPSS 13.0 was used to two groups to do t test for statistical analysis in test score and x2 test for degree of satisfaction.Results In the final examination, the score of non-case test of combinational teaching group was similar to that of traditional teaching group (50.30 ± 7.19 vs.52.04 ± 8.01, P=0.358).The combinational teaching group had significant improvement in case analysis test (35.76 ± 5.28 vs.31.80 ± 5.16), and the difference was statistically significant (P=-0.003).In the course of teaching satisfaction survey, the dual track teaching group, compared with the conventional teaching group, has a better effect on self study ability, communication skills, communication skills, and higher satisfaction for teaching, and more willing to continue to carry out teaching (P＜0.05).Conclusion The PBL+CBL combinational teaching model can make a great contribution to improving the teaching quality and satisfaction, and worthy of being popularized and applied.
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Based on the characteristics of oncology clinical teaching and the defects of current teaching methods,we launched teaching reform including the combination of basic and clinical medical curriculum,the practice of clinical problem-based learning (CPBL) method and the introduction of some guidance for medical practice.The teaching reform was effective,but there were drawbacks for improvement.
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This study examined the radiation-induced ERBB2 nuclear transport in the BT474 breast cancer cell line and the relationship between caveolin-1 and radiation-induced ERBB2 nuclear transport. The BT474 cells were treated with herceptin (200 nmol/L), PP2 (a caveolin-1 inhibitor, 100 nmol/L) and irradiation combined or alone. Confocal microscopy was used to observe the nuclear import of ERBB2 and caveolin-1 after irradiation. Western blotting was employed to detect the expression of ERBB2, caveolin-1 and DNA-PKcs after irradiation, and immunoprecipitation to identify the ERBB2 and caveolin-1 complex before perinuclear ERBB2 localization. Confocal microscopy showed the transport of ERBB2 and caveolin-1 from the cell membrane to the nucleus 15 min after irradiation and the proteins accumulated at the perinuclear region within 45 min. Western blotting revealed that the expression levels of ERBB2, caveolin-1 and DNA-PKcs were increased after irradiation and reached a peak 45 min later. Both herceptin and PP2 treatments were found to decrease ERBB2 expression. An immune complex composed of ERBB2 and caveolin-1 was found in the herceptin group after irradiation. It was concluded that after irradiation, ERBB2 may be transported from the cell membrane to the nucleus and activate DNA-PKcs to trigger DNA double-strand break (DSB) repair; caveolin-1 may participate in this process. Treatments involving the downregulation of caveolin-1 may increase the radiosensitization of breast cancer cells.
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Chemotherapy-induced neuropathy is a serious clinical problem for patients receiving cancer treatment. The aim of this study was to investigate the potential efficacy of pregabalin in chemotherapy-induced neuropathy in rats. A total of 35 male Sprague-Dawley rats were randomly divided into 5 groups: group 1, naive control; group 2, treated with pregabalin (30 mg/kg p.o., for 8 days); group 3, docetaxel was given by single intravenous infusion at 10 mg/kg; groups 4 and 5, pregabalin at 10 mg/kg and 30 mg/kg respectively was orally administered for 8 days after the docetaxel treatment. On day 8, behavioral test was performed, and substance P and CGRP release in dorsal root ganglion (DRG) and sciatic nerve were analyzed by electron microscope. Our results showed that docetaxel induced mechanical allodynia, mechanical hyperalgesia, heat hypoalgesia, cold allodynia, and sciatic nerve impairment and substance P and CGRP release in DRG. However, oral administration of pregabalin (10 mg/kg and 30 mg/kg) for 8 consecutive days significantly attenuated docetaxel-induced neuropathy by ameliorating heat hypoalgesia, cold allodynia, impairment of sciatic nerve and reducing the release of substance P and CGRP. The findings in the present study reveal that pregabalin may be a potential treatment agent against chemotherapy-induced neuropathy.
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This study examined the radiation-induced ERBB2 nuclear transport in the BT474 breast cancer cell line and the relationship between caveolin-1 and radiation-induced ERBB2 nuclear transport. The BT474 cells were treated with herceptin (200 nmol/L), PP2 (a caveolin-1 inhibitor, 100 nmol/L) and irradiation combined or alone. Confocal microscopy was used to observe the nuclear import of ERBB2 and caveolin-1 after irradiation. Western blotting was employed to detect the expression of ERBB2, caveolin-1 and DNA-PKcs after irradiation, and immunoprecipitation to identify the ERBB2 and caveolin-1 complex before perinuclear ERBB2 localization. Confocal microscopy showed the transport of ERBB2 and caveolin-1 from the cell membrane to the nucleus 15 min after irradiation and the proteins accumulated at the perinuclear region within 45 min. Western blotting revealed that the expression levels of ERBB2, caveolin-1 and DNA-PKcs were increased after irradiation and reached a peak 45 min later. Both herceptin and PP2 treatments were found to decrease ERBB2 expression. An immune complex composed of ERBB2 and caveolin-1 was found in the herceptin group after irradiation. It was concluded that after irradiation, ERBB2 may be transported from the cell membrane to the nucleus and activate DNA-PKcs to trigger DNA double-strand break (DSB) repair; caveolin-1 may participate in this process. Treatments involving the downregulation of caveolin-1 may increase the radiosensitization of breast cancer cells.
Subject(s)
Female , Humans , Active Transport, Cell Nucleus , Physiology , Breast Neoplasms , Metabolism , Caveolin 1 , Metabolism , Cell Line, Tumor , Protein Transport , Physiology , Radiation , Receptor, ErbB-2 , MetabolismABSTRACT
Chemotherapy-induced neuropathy is a serious clinical problem for patients receiving cancer treatment. The aim of this study was to investigate the potential efficacy of pregabalin in chemotherapy-induced neuropathy in rats. A total of 35 male Sprague-Dawley rats were randomly divided into 5 groups: group 1, naive control; group 2, treated with pregabalin (30 mg/kg p.o., for 8 days); group 3, docetaxel was given by single intravenous infusion at 10 mg/kg; groups 4 and 5, pregabalin at 10 mg/kg and 30 mg/kg respectively was orally administered for 8 days after the docetaxel treatment. On day 8, behavioral test was performed, and substance P and CGRP release in dorsal root ganglion (DRG) and sciatic nerve were analyzed by electron microscope. Our results showed that docetaxel induced mechanical allodynia, mechanical hyperalgesia, heat hypoalgesia, cold allodynia, and sciatic nerve impairment and substance P and CGRP release in DRG. However, oral administration of pregabalin (10 mg/kg and 30 mg/kg) for 8 consecutive days significantly attenuated docetaxel-induced neuropathy by ameliorating heat hypoalgesia, cold allodynia, impairment of sciatic nerve and reducing the release of substance P and CGRP. The findings in the present study reveal that pregabalin may be a potential treatment agent against chemotherapy-induced neuropathy.
Subject(s)
Animals , Male , Rats , Ganglia, Spinal , Nervous System Diseases , Drug Therapy , Pregabalin , Rats, Sprague-Dawley , Sciatic Nerve , Taxoids , gamma-Aminobutyric Acid , PharmacologyABSTRACT
Objective To observe the influence of trastuzumab on DNA break repair and Her-2 nuclear import after radiation in breast cancer cell line SKBR3,and discuss the radiosensitivity mechanism of trastuzumab.Methods Clone formation assay was used to analyze the difference of survival fractions between radiation group and radiation plus trastuzumab group.Confocal microscopy was applied to observe the influence of trastuzumab in the nuclear import process of Her-2 and the expression of γH2AX after radiation,which is considered as the marker of DNA double strand break.Western blotting was used to detect the expression of Her-2 and DNA-PKcs in nuclei after radiation.Results The result of clone formation assayshowed that the SF2 in radiation group was 0.547±0.046 and 0.321±0.022 in the radiation plus trastuzumab group were significantly decreased,the results of confocal microscopy showed that trastuzumab postponed the nuclear import process of Her-2 (52.80±19.74 in radiation group,21.41±10.55 in the radiation group),and increased expression of γH2AX after radiation (85.40±25.63 in radiation group,18.53±44.32 in the radiation group),and western blotting revealed trastuzumab reduced the expression of Her-2,DNA-PKcs in nuclei.Conclusion Trastuzumab can inhibit the radiation induced nuclear import of Her-2,and decrease Her-2,DNA-PKcs in nuclei to increase the DSB on early stage after radiation.
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Hypermethylation in the promoter region of tumor suppressor genes is a common mechanism of gene silencing, which tends to occur in cancer. The effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR), a specific DNA methyltransferase inhibitor, on the cell proliferation of human breast cancer cell line MCF-7 and on the expression of Apaf-1 gene were investigated. Human MCF-7 cells were incubated with increasing concentrations of 5-Aza-CdR for 12 to 120 h. The growth inhibition rates of MCF-7 cells were detected by MTT assay. Changes of cell cycle distribution and apoptotic rates of MCF-7 cells were determined by flow cytometry. The expressions of DNA methyltransferase 3b mRNA and Apaf-1 mRNA were measured by reverse transcription polymerase chain reaction (RT-PCR). Meanwhile, the expression of Apaf-1 protein was detected by Western blotting. The results showed that 5-Aza-CdR significantly inhibited the growth of MCF-7 cells and the growth inhibition rate of MCF-7 cells was significantly enhanced with the concentration of 5-Aza-CdR and the action time. Flow cytometry indicated that 5-Aza-CdR could significantly induce G(1)/S cell cycle arrest and increase the apoptosis rate of MCF-7 cells. The mRNA and protein expressions of Apaf-1 were up-regulated in MCF-7 cells treated with 5-Aza-CdR, which was accompanied by down-regulation of DNA methyltransferase 3b mRNA. It is concluded that 5-Aza-CdR might retard the growth of tumor cells and promote the apoptosis of MCF-7 breast cancer cells by inhibiting the expression of DNA methyltransferase 3b and re-activating the Apaf-1 gene expression.
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Objective To evaluate the influence of belly beard device and the distended bladder on the dose distribution of PTV and the dose-volume histograms(DVHs)of organs at risk(OARs)for postoperative radiation tIlerapy of rectal cancer.Methods A total of 23 patients(8 and 15)with distended bladder receiving 3-field postoperative radiation therapy were dealed with or without a special belly beard in the prone position.At the same time,15 cages with belly board were scanned with empty bladder.The volume of irradiated small bowel was calculated for doses between 5-50 Gy at 5 Gy intervals.With prescription dose in plan target volume(PTV)of 50 Gy,we compared the dose distribution,DVH of OARs,conformity index(CIPTV),the volume of irradiated small bowel and the acute toxicity under the condition of thlee different moulds.Results There was no significant difference in PTV's converge,DVHs of femoral head and CI among 3 moulds(P＞0.05).With the belly board,the high-dose volume of irradiated small bowel(V20-V52.5)was significantly decreased(P＜0.05),specially with distended bladder.However,the low dose volume(V5-V15)was slightly increased.The bladder distension significanfly decreases the volumes of the irradiated small howel at dose levels from 15-52.5 Gy(P＜0.05).Furthermore,the mean volume(V5-V30)of irradiated small bowel differed significantly between patients experiencing Grade 0.1 and ≥2 diarrhea(P＜0.05).Conclusions The combination of belly board and distended bladder was more effectively to reduce the irradiated small bowel volume among 3 moulds,so as to minimized acute diarrhea toxicity.
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Objective To observe the EGFR nuclear translocation in cervical carcinoma cell lines after irradiation and its possible role in radiation tolerance.Methods Western blotting was used to detect the nuclear EGFR and cytoplastic EGFR after irradiation.The effect of Cetuximab on expression of nuclear EGFR and survival fractions were investigated.Results After irradiation,compared with control group,the expression of nuclear EGFR protein increased in irradiated cervical carcinoma cell.Cetuximab inhibited the radiation-induced nuclear EGFR expression with decreased survival fractions.Conclusion Radiation could induce EGFR nuclear translocation in cervical carcinoma cell lines and nuclear EGFR might be correlated with radiation tolerance in Cervical carcinoma cell.
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<p><b>BACKGROUND</b>Survivin is a rather specific gene in tumor tissue. We transfected dendritic cells (DCs) with recombinant adenovirus (Ad) containing survivin gene and granulocyte-macrophage colony-stimulating factor (GM-CSF) gene and tested the inducing effect of the transfected DCs on cytotoxic T lymphocytes (CTL) to kill leukemic cells.</p><p><b>METHODS</b>After derived from the peripheral, DCs was assayed by mixed leukocyte reaction (MLR) tests. Lactate dehydrogenase (LDH) release test was used to evaluate cytotoxicity of CTL.</p><p><b>RESULTS</b>Expression of survivin in transfected DCs was confirmed by Western blotting analysis. GM-CSF expression was confirmed by enzyme-linked immunosorbent assay (ELISA). In MLR assay, DCs coinfected with Ad-survivin and Ad-GM-CSF induced higher allogeneic lymphocyte reaction than control DCs at ratios of 1:5, 1:10, 1:50 and 1:100. DCs coinfected with Ad-survivin and Ad-GM-CSF had much higher activity of CTL to HL-60 cells than DCs infected with Ad-survivin only, Ad-GM-CSF only, or control DCs. Levels of interleukin-12 (IL-12) and interferon gamma (IFN-gamma) in lymphocyte supernatants containing DCs coinfected with Ad-survivin and Ad-GM-CSF were significantly higher than those in the control group.</p><p><b>CONCLUSION</b>DCs coinfected with Ad-survivin and Ad-GM-CSF induce much higher anti-leukemic response in vitro than those infected with either factor. Therefore, adenovirus vectors containing survivin and GM-CSF genes may be promising vaccine candidates for leukemia therapy.</p>
Subject(s)
Humans , Adenoviridae , Genetics , Cytotoxicity, Immunologic , Dendritic Cells , Physiology , Genetic Therapy , Granulocyte-Macrophage Colony-Stimulating Factor , Genetics , HL-60 Cells , Inhibitor of Apoptosis Proteins , Interferon-gamma , Interleukin-12 , Leukemia , Therapeutics , Lymphocyte Activation , Microtubule-Associated Proteins , Genetics , Neoplasm Proteins , Genetics , T-Lymphocytes, Cytotoxic , Allergy and Immunology , TransfectionABSTRACT
Objective: To study the changes in radiosensitivity and cell cycle distribution of HeLa cells after inhibition of one or several DNA double-strand break (DSB) repair proteins by small interfering RNA (siRNA) and LY294002. Methods: Ku80 silenced cells (HeLa/Ku8O-siRNA) and control cells (HeLa/ Neg-siRNA) were transfected with siRNA targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) or pretreated with 50 μmol/L LY294002, a chemically specific inhibitor of DNA-PKcs. After the cells received 6MV X-ray irradiation, the radiosensitivity of the cells was detected by clony formation assay and cell cycle distribution was analyzed by flow cytometry. Results: The survival fraction at 2 Gy (SF2) value was 0.08 ± 0.01 for HeLa/Ku80-siRNA cells after being transfected with DNA-PKcs siRNA; the SF2 value reached 0.03 ± 0.01 when HeLa/Ku80-siRNA cells were pretreated with LY294002. Both of them were significantly lower than that of untreated HeLa/Ku80-siRNA cells (0.20 ± 0.05). The cells in all the groups were arrested in G2/M phase after irradiaton with 6 Gy X-ray. The G2/M arrest occurred slowly in the DNA -PKcs-siRNA-transfec ted HeLa/Neg-siRNA cells and LY294002-pretreated HeLa/Ku80-siRNA and HeLa/Neg-siRNA cells, which did not reach the peak at 72 h post-irradiation. The G2/M accumulation was maximal at 48 h post-irradiation in other cell lines. Conclusion: Based on 95% inhibition of Ku80 protein, DNA-PKcs or ataxia-telangiectasia mutant (ATM) gene could compensate the DSB repair function. Co-inhibition of these proteins led to increase in radiosensitivity of HeLa cells; Ku80, DNA-PKcs and ATM play different roles in G2/M arrest.
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Objective: To explore the correlation between the expressions of DSB (DNA double-strand break) repair protein (including Ku80, DNA-PKcs, and ATM) and radiosensitivity parameters of human tumor cell lines, and to reveal the value of the three proteins for the prognosis of the radiosensitivity of tumor cells. Methods: Eight tumor cell lines were selected including four human cervical carcinoma cell lines (HeLa, SiHa, C33A, and Caski), three human breast carcinoma cell lines (MCF-7, MDA-MB-231, MDA-MB453), and one human lung carcinoma cell line (A549). The expressions of Ku80, DNA-PKcs and ATM protein were measured by Western blotting. The apoptotic ratio of tumor cells was analyzed by flow cytometry after 48 h X-ray irradiation at 10 Gy of 6 MV. SF2 value (survival fraction at 2 Gy) and α and β values were obtained by clone formation assay. The correlation of protein expression with SF2, α/β value or apoptotic ratio was analyzed by Pearson linear correlation analysis. Results: The expression of same protein in different cell lines and the expression of the three proteins in the same cell line had significant difference. There was a positive correlation between the expression of DNA-PKcs and SF2 (r=0.723, P=0.043 0.05). The expression of the three proteins had no correlation with either apoptotic ratio or α/ β value (P>0.05). Conclusions: Tumor cells with higher expression of DNA-PKcs protein will have higher radioresistance. The expression level of DNA-PKcs protein in tumor cells may be an indicator for predicting the radiosensitivity of tumor cells.
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Objective To investigate the significance of the serum levels of interleukin-10 (IL-10),IL-13,IL-15 of patients with chronic hepatic failure and the correlation between those inter- leukin levels and nosocomial infections.Methods The serum levels of IL-10,IL-13,IL-15 of 58 patients with chronic hepatic failure were measured by double antibody sandwich enzyme-linked immu- nosorbent assay at the time of admission and 2 weeks after admission.Results The serum levels of IL-15 and the propotion of IL-15/IL-10 and IL-15/IL-13 in patients with chronic hepatic failure group at the time of admission were significantly higher than those in healthy control group[(358.16?290.91) ng/L vs (38.55?21.49) ng/L,12.93?14.26 vs 1.10?0.55,98.55?97.5.5 vs 9.70?5.03,respectively,all P=0.000].Those in death group were significantly higher than those in improving group[(479.93v205.52) ng/L vs (244.51?236.29) ng/L,17.65?17.78 vs 8.53?7.98,130.69?115.50 vs 68.55?65.99,respectively,all P
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In order to investigate the role of antiapoptosis gene, survivin in the resistance to palcitaxel, the expression of survivin mRNA and protein in the process of paclitaxel treatment in breast cancer cell line MCF-7 was detected MCF-7 cells were incubated with paclitaxel at different concentrations. The growth inhibition rate of MCF-7 was investigated by tetrazolium bromide (MTT) colorimetry. The change of apoptosis was detected by Annexin-V/PI methods. The changes in the expression of survivin mRNA and protein were studied by reverse transcription polymerase chain reaction (RT-PCR) and Western-blot assay respectively. The growth inhibition rate of MCF-7 was increased in a concentration-and time-dependent manner. Paclitaxel of higher concentration could effectively induce apoptosis in MCF-7 cells after 48 h, while the expression of survivin was increased at early time (within 6 h) and decreased after 24 h regardless of treatment concentrations of paclitaxel. It suggested that tumor cells might evade the paclitaxel-induced cell cycle arrest and apoptosis by increasing the level of survivin at early treatment time.
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In order to investigate the role of antiapoptosis gene, survivin in the resistance to palcitaxel, the expression of survivin mRNA and protein in the process of paclitaxel treatment in breast cancer cell line MCF-7 was detected. MCF-7 cells were incubated with paclitaxel at different concentrations. The growth inhibition rate of MCF-7 was investigated by tetrazolium bromide (MTT) colorimetry. The change of apoptosis was detected by Annexin-V/PI methods. The changes in the expression of survivin mRNA and protein were studied by reverse transcription polymerase chain reaction (RT-PCR) and Western-blot assay respectively. The growth inhibition rate of MCF-7 was increased in a concentration- and time-dependent manner. Paclitaxel of higher concentration could effectively induce apoptosis in MCF-7 cells after 48 h, while the expression of survivin was increased at early time (within 6 h) and decreased after 24 h regardless of treatment concentrations of paclitaxel. It suggested that tumor cells might evade the paclitaxel-induced cell cycle arrest and apoptosis by increasing the level of survivin at early treatment time.
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In order to investigate the effects of short hairpin RNA (shRNA) on the expression of Survivin, cell cycle and cell proliferation in MCF-7 cells, using a pEGFP vector which contained a U6 promoter shRNA plasmid targeted against survivin was constructed and transfected into MCF-7 cells. The change of the expression of Survivin and cell proliferation rates were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and MTT methods respectively. The change of cell cycle after transfection was analyzed by flow cytometry. The results indicated that the recombinant plasmid containing Survivin shRNA was constructed successfully, which could suppress the expression of Survivin at mRNA and protein level. The growth of MCF-7 cells was arrested in G1 phase of the cell cycle and the proliferation activity was suppressed after transfection. It was concluded that Survivin shRNA plasmid could knock down the expression of Survivin in MCF-7 cells specifically. In addition, Survivin shRNA plasmid could lead to G1 arrest and inhibit the proliferation of MCF-7 cells, which suggested that Survivin shRNA might be used as a new therapeutic method for breast cancer.
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In order to investigate the effects of short hairpin RNA (shRNA) on the expression of Survivin, cell cycle and cell proliferation in MCF-7 cells, using a pEGFP vector which contained a U6 promoter shRNA plasmid targeted against survivin was constructed and transfected into MCF-7 cells. The change of the expression of Survivin and cell proliferation rates were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and MTT methods respectively. The change of cell cycle after transfection was analyzed by flow cytometry. The results indicated that the recombinant plasmid containing Survivin shRNA was constructed successfully, which could suppress the expression of Survivin at mRNA and protein level. The growth of MCF-7 cells was arrested in G1 phase of the cell cycle and the proliferation activity was suppressed after transfection. It was concluded that Survivin shRNA plasmid could knock down the expression of Survivin in MCF-7 cells specifically. In addition, Survivin shRNA plasmid could lead to G1 arrest and inhibit the proliferation of MCF-7 cells, which suggested that Survivin shRNA might be used as a new therapeutic method for breast cancer.