1.Expression and function of BRAF-activated long non-coding RNA in colorectal cancer
Qinhao GUO ; Yan ZHAO ; Jiejing CHEN ; Jun HU ; Shuwei WANG ; Dongsheng ZHANG ; Yueming SUN
Chinese Journal of Digestive Surgery 2014;13(5):376-380
Objective To investigate the expression of BRAF-activated long non-coding RNA (BANCR) in colorectal cancer,and the influence of BANCR on the biological function of HCT116 cells.Methods Fifty-six samples of colorectal cancer specimen (including the cancer tissues and precancerous tissues) were obtained at the First Affiliated Hospital of Nanjing Medical University from March 2012 to June 2013.The expressions of BANCR in all the specimens were detected by qRT-PCR (28 cases in the BANCR-high expression group and 28 cases in the BANCR-low expression group).The relationship between the expressions of BANCR and the clinicopathological factors of colorectal cancer was analyzed.The HCT116 cells were divided into 4 groups after interfering BANCR with lentiviral-mediated shRNA-1 and shRNA-2:interference group 1 (HCT116 cells transfected with LV-shRNA-1),interference group 2 (HCT116 cells transfected with LV-shRNA-2),negative control group (HCT116 cells transfected with lentivirus vector with nonsense sequence) and blank control group (HCT116 cells cultured in RPMI 1640 medium).The proliferation,apoptosis and migration of HCT116 cells in the 4 groups were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively.The comparison between the 2 groups was analyzed by u test,and multiple groups were compared by one-way analysis of variance,repeated measurement analysis of variance and LSD-t test.Multivariate analysis was done by Logistic regression model.The difference between categorical data was compared by chi-square test.Results The relative expression of BANCR in the cancer tissues was 1.6 ± 0.4,which was significantly higher than 0.9 ± 0.7 of the precancerous tissues (u =1 020.000,P < 0.05).The result of univariate analysis showed that the high expression of BANCR was correlated with the lymph node metastasis and tumor stage (x2 =4.595,7.487,P < 0.05).The result of multivariate analysis showed that lymph node metastasis and tumor stage (stage Ⅲ-Ⅳ) were the independent risk factors influencing the high expression of BANCR(OR =4.000,5.914,95% CI:1.230-12.900,1.685-20.760,P < 0.05).The relative expressions of BANCR of the interference group 1,interference group 2,negative control group and the blank control group were 0.25 ±0.04,0.20±0.06,0.96 ±0.04,0.98 ±0.03,with significant difference among the 4 groups (F =271.610,P < 0.05).The cell proliferation rates at day 6 of the interference group 1,interference group 2 and the negative control group were 80.6% ± 7.6%,81.2% ± 5.1% and 87.9% ± 13.6%,with no significant difference among the 3 groups (F =0.559,P > 0.05).The apoptotic rates of the interference group 1,interference group 2,negative control group and the blank control group were 4.7% ± 1.7%,5.1% ± 1.1%,3.1% ± 0.6% and 2.8% ± 0.9%,with no significant difference among the 4 groups (F =2.881,P > 0.05).The numbers of transmembrane cells of the interference group 1,interference group 2,negative control group and the blank control group were 135 ± 29,107 ± 18,240 ± 24 and 245 ± 22,with significant difference among the 4 groups (F =45.194,P < 0.05).Conclusions BANCR was overexpressed in the HCT116 cells,and the BANCR overexpression was correlated with lymph node metastasis and tumor stage.BANCR can promote the migration of HCT116 cells.BANCR could be an important biomarker for the diagnosis and prognosis of colorectal cancer.
2.First trimester combined screening for Down's Syndrome with NT, Free-βhCG and PAPP-A
Zhengyou MIAO ; Tongkun SHI ; Yanjun GUO ; Qinhao SONG ; Huaxiang SHEN ; Ying XU
Chinese Journal of Laboratory Medicine 2012;35(10):932-935
Objective To explore the sensitivity of using NT,combined with serum biochemical markers (Free-βhCG,PAPP-A) for Down's Syndrome screening in early stage of pregnancy.Methods Collect pregnant women aged 17-45 years old voluntary antenatal screening in our hospital from March 2009to October 2010,a total of 11882 cases.Serum Free-βhCG and PAPP-A were measured NT value was determined by ultrasound at 11-13w+64 of gestation.Calculating combined screening (NT,Free-βhCG,PAPP-A),and serum integrated screening (Free-beta hCG,PAPP-A) risk,respectively,using the risk calculation software for the same person.Results Early pregnancy screening was performed in 11 882patients,18 had a fetus with Down's syndrome,a rate of 0.15%.The detection rates of Down's syndrome in combined screening and serum integrated screening were 83.3% and 72.2% respectively.The specificities were 98.4% and 97.3% and detection efficiency were 7.18%,3.90% respectively.Areas under the curve (AUCs) of fhst-trimester combined screening and serum integrated screening were 0.975 (95% CI:0.943,1.007),0.901 (95% CI:0.789,1.013) respectively.Conclusion In early stage of pregnancy,combined screening for Down's syndrome has higher sensitivity and specificity than serological screening,has higher detection rate in the same false-positive rate case,which can effectively reduce the pregnant women to receive invasive puncture.
3.Multiple of the median correction of Down's syndrome serum markers with pregnant body weight
Zhengyou MIAO ; Yanjun GUO ; Tongkun SHI ; Qinhao SONG ; Xia LIU ; Ying XU
Chinese Journal of Laboratory Medicine 2014;37(1):46-49
Objective With Pregnant women weight correction for serum marker Multiple of Median (MoM) of First trimester and second-trimester,integrated screen-ing for Down's syndrome (DS),can reduce the false positive rate.Methods The same pregnant woman were taken venous blood vessels with sterile vacuum during the first trimester (11-13 W(+ 6) d) and the second trimester (15-20 W(+ 6) d),Alpha-fetoprotein (AFP),serum free beta-human chorionic gonadotrophin (Free beta hCG) and pregnancyassociated protein-A (PAPP-A) of three kinds of serum marker screening indicators were assayed by Using Time-resolved fluoroimmunoassay (TRFIA).Screening for risk assessment software was used to calculate serum marker Multiple of Median,To assess the risks of 7 997 cases of local pregnant women DS,To construct the weight equation of local population using nonlinear weighted regression method,With maternal weight correction for serum marker Multiple of Median (MoM) of local pregnant women,Comparing the changes of screening index MoM before and after correction,chi square test to compare the detection rate and false positive rate.Results MoM values of three kinds of serum markers (al-pha-fetoprotein,free beta subunit of human chorionic gonadotropin,pregnan-cy-associated plasma protein A) decreased with the weight increasing,Screening index MoM after correction weight equation,the screening of false positives for crowd from 4.12% down to 3.86% (x2 =0.021,P > 0.05).Setting threshold (cut-off) at 1/270,and no change detection rates were 71.4% the local population before and after correction weight equation.Conclusion Maternal weight may affect the results of Down's syn-drome sereening.When screening proposal to set up,it is worth making weight cor-rections for serum maker multiple of median in order to get accurate risk calculation results.