miRNA-141通过抑制MAP2K4促进大肠癌增殖的研究

目的:探究miRNA-141通过抑制MAP2K4对促进大肠癌增殖的效果。方法:本文共选取2016年2月至2018年2月在本院放射科做MRI检查被诊断为大肠癌的患者24例,提取总RNA,RNA浓度测量使用紫外分光光度法,取5μL RNA样品,运用RNase对离子水进行稀释,在对OD值进行测量时需使用紫外分光光度计进行测量。RT-PCR扩增,cDNA模版的合成主要是应用PCR逆转录反应来实现,RT-PCR扩增主要是应用ALL-in-One试剂盒,具体的步骤及方法应严格按照试剂盒上的说明书开展。结果:大肠癌及肠黏膜的提取方法主要是应用Trizol方法。在对miR-141的相对表达量进行检测时,主要是采用荧光实时定量PCR法。在对细胞转染WT FL质粒进行检验时,主要是使用荧光仪来检测荧光酶的活性,会发现FL活性下降,突变质粒的FL活性未发生明显的变化。结论:在大肠癌诊断中MRI发挥了重要作用,miR-141的表达水平会随着大肠癌TNM分期而增高,miR-141表达上调,说明大肠癌与miR-141之间的发展有直接关系,有助于抑制癌细胞的快速生长。

Effect of preoperative FOLFOX chemotherapy on CCL20/CCR6 expression in colorectal liver metastases

AIM： To evaluate the influence of preoperative FOLFOX chemotherapy on CCL20/CCR6 expression in liver metastases of stage IV colorectal cancer （CRC） patients.（/7 = 53） and in patients who did not receive FOLFOX chemotherapy prior to liver surgery （n = 29）. RESULTS： Of the 53 patients who received FOLFOX, time to liver surgery was n〈 1 mo in 14 patients, 〈 1 year in 22 patients and 〉 1 year in 17 patients, respectively. In addition, we investigated the proliferation rate of CRC cells in liver metastases in the different patient groups. Both CCL20 and CCR6 mRNA and protein ex- pression levels were significantly increased in patients who received preoperative FOLFOX chemotherapy ~〈 12 mo before liver surgery （P 〈 0.001） in comparison to patients who did not undergo FOLFOX treatment. Further, proliferation of CRLM cells as measured by Ki-67 was increased in patients who underwent FOLFOX treat- ment. CCL20 and CCR6 expression levels were significantly increased in CRLM patients who had undergone preoperative FOLFOX chemotherapy. CONCLUSION： This chemokine/receptor up-regulation could lead to increased proliferation/migration through an autocrine mechanism which might be used by surviving metastatic cells to escape cell death caused by FOLFOX.

Influence of Deleted in Colorectal Carcinoma Gene on Proliferation of Ovarian Cancer Cell Line SKOV-3 In Vivo and In Vitro

Objective To elucidate the effects of the deleted in colorectal carcinoma(DCC) gene on proliferation of ovarian cancer cell line SKOV-3.Method An exogenous recombinant eukaryotic expression vector pcDNA3.1(+)-DCC,containing human DCC cDNA coding sequences,was constructed and transfected into SKOV-3 cells(SKOV-3/DCC).The pcDNA3.1(+) transfected cells(SKOV-3/Neo) and SKOV-3 cells were used as the positive and negative controls,respectively.Expressions of DCC mRNA and protein were analyzed by RT-PCR and immunocytochemical analysis,respectively.Cell growth was detected by soft agar colony formation assay and MTT assay.Flow cytometry and transmission electron microscopy were used to assess the effects of DCC on cell cycle distribution and ultrastructure,respectively.BALB/c mice were used to evaluate the effects of DCC on tumorigenicity in vivo.Results RT-PCR and immunocytochemical analysis revealed the exogenous DCC gene was successfully transfected into SKOV-3 cell lines and obtained permanent expression.The half maximal inhibitory concentration(IC50) of SKOV-3/DCC cells was significantly lower than that of SKOV-3 or SKOV-3/Neo cells(all P<0.05).DCC expression caused SKOV-3 cells to be arrested in G1 phase(78.0%),and electron microscopic analysis showed SKOV-3/DCC cells displayed typical morphological changes of apoptosis.Two mice xenografted with SKOV-3/DCC cells showed no tumor tumorigenecity.The tumor volume of BALB/c mice bearing SKOV-3/DCC cells(3.403 mm3) was smaller than that of SKOV-3 cells(9.206 mm3).Conclusion DCC gene may play an important role in suppressing the growth of SKOV-3 cell line and inducing apoptosis.

Pin1 expression affects cell proliferation and apoptosis of SW620 cells in colorectal carcinoma

Objective:The aim of our study was to investigate the effects of Pin1 reduction on SW620 cell proliferation and apoptosis in human colorectal carcinoma.Methods:We constructed a plasmid of RNA interfering(shRNA) for Pin1 gene(pGenesl-1-Pin1),then the plasmid was transfected into colorectal carcinoma SW620 cells line by liposome mediation.The protein expression of Pin1 was tested by Western blotting.The proliferation rate was analyzed by MTT and the apoptotic rate of cells was tested by flow cytometry.In order to explain further the effect of Pin1 in SW620 cells,the protein level of Bcl-2 was analyzed by Western blotting.Results:pGenesil-1-Pin1 plasmid was successfully constructed and confirmed by sequencing.The protein relative levels of Pin1 were 0.06 ± 0.04 for the P-shRNA/SW620 cells,and 0.32 ± 0.09 for the P-Con/SW620 cells.The cell growth rate of SW620 cells was slower while the apoptotic rate was increased after transfection with pGenesil-Pin1 plasmid,and the apoptotic rate was 12.38% ± 1.55% for the P-shRNA/SW620 group.At the same time,we found that the protein expression of Bcl-2 was also reduced.The results were 0.13 ± 0.04 for the P-shRNA/SW620 cells,and 0.36 ± 0.08 for the P-Con/SW620 cells.Conclusion:Inhibited Pin1 expression may suppress the cell proliferation and promote apoptosis of colorectal carcinoma cells in vitro.