1.The expression and significance of MDR1 ,BCRP,LRP mRNA and protein in breast cancer
Jie CUI ; Xinlan LIU ; Ningju WANG
Chinese Journal of Laboratory Medicine 2010;33(12):1164-1170
Objective To investigate the expression and the significance of the MDR1, breast cancer resistance protein, lung cancer resistance protein mRNA and corresponding proteins P-gp, BCRP and LRP in breast cancer tissues and adjacent breast tissues. Methods RT-PCR was used to exam the expression of MDR1, BCRP, LRP mRNA in 42 breast cancer tissues and 42 adjacent tissues. IHC was used to exam the expression of P-gp, BCRP and LRP in 126 breast cancer tissues and 42 adjacent tissues, and theirs relationships with clinicopathological parameters in breast cancer, axillary lymph node status and 5-year recurrence and metastasis. Results The relative expression levels of MDR1, BCRP and LRP mRNA were 0.81 ±0.17,0.78 ±0.14,0.79 ±0.13 in breast cancer tissues and 0.33 ±0.11,0.45 ±0.09,0.36 ±0.10 in adjacent tissues respectively. There were significant differences between cancer tissues and adjacent tissues ( t = 4.613, 4.850 and 8. 089, P < 0.01 ). The positivities of P-gp, BCRP and LRP were 41% ( 52/126) ,39% (49/126) and 66% (83/126) in breast cancer tissues. There were significant differences between cancer tissues and adjacent tissues (x2 = 10.147, 7.020, 27.820, P < 0.01 ). The expression of MDR1 mRNA/P-gp was significantly associated with tumor stage and lymph node metastasis ( r = 0.369,0.398, P < 0.05 ). The expression of BCRP (mRNA/protein) was significantly associated with lymph node metastasis (r = 0.355, P < 0.05 ) . The positivities of P-gp were significantly different between 39 recurrence/metastasis patients occurred in 5 years and 32 unrecurrence/nonmetastasis patients in 5 years (x2 = 11.771, P < 0.01 ). The positivities of BCRP and LRP were not significantly different in these two groups(x2 =2.261,0.078,P >0.05). The coincidence rates for expression of MDR1 ,BCRP,LRP mRNA and their proteins in breast cancer tissues were 90.48% ,92.85% and 85.71% respectively (the Kappa values were 0.806,0.751 and 0.697, P < 0.01 ). Conclusions Multidrug resistance is common in breast cancer. The three drug resistance genes and proteins are involved in the formation of multidrug resistance of breast cancer. Detection of multidrng resistance genes in breast cancer may be useful to choose chemotherapy,especially patients with P-gp positive expression are not advised to use the CAF chemotherapy.
2.Effects of VEGF-C, VEGFR-3 and LYVE-1 on Lymphangiogenesis and Prognosis in Gastric Cancer
Xinlan LIU ; Yanjiao ZHAO ; Zhilan MA
Tianjin Medical Journal 2013;(8):747-751
Objective To study the role of expressions of vascular endothelial growth factor C (VEGF-C), vascular endothelial growth factor receptor-3 (VEGFR-3) and lymphatic vessel hyaluronan receptor-1(LYVE-1) on the lymphangio-genesis and prognosis in gastric cancer. Methods The tissue microarray technology was used to detect the expressions of VEGF-C, VEGFR-3 and LYVE-1 in 125 gastric cancer specimens, 96 adjacent normal tissues and 20 benign gastric lesion samples. The lymphatic vessel density (LVD) marked by Podoplanin was detected as well. Results The positive rates of VEGF-C, VEGFR-3 and LYVE-1 in gastric cancer tissues were 62.4%, 56.0%and 58.4%, which were significantly higher than those in adjacent normal tissues (10.4%,12.5%and 9.4%) and benign gastric lesion tissues (20%, 30%and 25%, P<0.05). The LVD score was significantly higher in gastric intra-tumoral and peri-tumoral samples (2.98±0.81 and 4.22±1.09) than that in adjacent normal tissues or benign gastric lesion samples (1.82±0.63 or 0.89±0.45, P<0.01). The LVD score was significantly higher in peri-tumoral samples than that of intra-tumoral samples (P<0.01). There was a positive relationship between expression levels of VEGF-C,VEGFR-3 and LYVE-1 with LVD (P<0.05). The positive expressions of the three indexes were the risk factors of lymph node metastasis and distant metastasis (P<0.05). There was a significantly longer 5-year survival rate in patients with negative expression of the three indexes (P<0.05). Conclusion VEGF-C, VEGFR-3 and LYVE-1 proteins were positively highly expressed in gastric cancer tissues, which were risk factors affecting the progno-sis of gastric cancer. The expression levels of the three indexes can be used to predict the prognosis and lymphangiogenesis of gastric cancer.
3.The correlation and significance of gene polymorphisms of C1236T, G2677T/A and C3435T with molecular subtypes of breast cancer
Xinlan LIU ; Haixia ZHANG ; Yaobang LIU ; Min JIANG
Tianjin Medical Journal 2016;44(4):389-393
Objective To investigate the distribution of the MDR1 exon12 (C1236T), exon21 (G2677T/A) and exon 26 (C3435T) gene polymorphisms in breast cancer patients, and to analyse their relationship with molecular subtypes of breast cancer. Methods The genotyping of C1236T, G2677T/A and C3435T were detected by polymerase chain reaction (PCR)-high resolution melting (HRM) method in 400 cases of breast cancer. The Hardy-Weinberg equilibrium test was used for ge?netic equilibrium distribution of genotype. The molecular subtypes of breast cancer were classified based on St.Gallen Con?sensus 2013. The genotype distributions of C1236T, G2677T/A and C3435T in breast cancer were analyzed. Their relation?ship with molecular subtypes in breast cancer was analyzed as well. Results ①In 400 cases of breast cancer, there were 2, 3 and 2 specimens did not get genotyping results in C1236T, G2677T/A and C3435T genotype detection. The CC, CT and TT genotypes of C1236T accounted for 16.08% (64/398), 44.22% (176/398) and 39.70% (176/398). GG, GT, GA, TT and AT genotypes of G2677T/A accounted for 16.62%(66/397), 44.33%(176/397), 7.05%(28/397), 27.46%(109/397) and 4.54%(18/397). CC, CT and TT genotypes of C3435T accounted for 21.11%(84/398), 56.03%(223/398) and 22.86%(91/398) re?spectively. Hardy-Weinberg genetic equilibrium testing showed that polymorphisms of C1236T, G2677T/A and C3435T had group representation (P<0.05).②Eleven cases of HER-2 (2+) were excluded because they were not verified by FISH de?tection when performed molecular subtype of breast cancer. Luminal A subtype accounted for 41.90%(163/389), Luminal B subtype accounted for 32.65%(127/389), HER-2 over-expression subtype accounted for 13.62%(53/389) and triple nega?tive subtype accounted for 11.83%(46/389).③CT/TT genotype frequency of C3435T was significantly higher in breast can?cer patients with Luminal A subtype than that in breast cancer patients with HER-2 over-expression subtype and triple neg?ative subtype (χ2=12.011, P=0.001;χ2=13.976, P<0.001), while there was no statistical difference in C1236T and G2677T/A gene polymorphism between different molecular subtypes of breast cancer (P>0.05). Conclusion C3435T gene polymor?phism can explain more accurately heterogeneity of breast cancer. CT/TT genotype in different molecular subtypes of breast cancer may be more sensitive to drug treatment.
4.Diagnostic value of MRI in pituitary stalk interruption syndrome
Lianggeng GONG ; Xinlan XIAO ; Liling XIE ; Yuanyuan LIU ; Ting SHU
Chinese Journal of Radiology 2008;42(7):706-708
Objective To study characteristic findings and the diagnostic value of MRI in pituitary stalk interruption syndrome (PSIS). Methods Twenty-one patients with PSIS were included. Small field of view (FOV) MR1 scanning and clinic hormone detection were performed in all patients. Moreover, fluid attenuated inversion recovery (FLAIR)T,1 WI with fat-suppression sequence was performed in 8 cases. The appearance on FLAIR T1 WI and T2 WI were recorded. The shape of pituitary stalk and antehypophysis, and the signal intensity of posthypophysis were analyzed simultaneously. Results Growth hormone deficiency (GHD) was confirmed by clinic hormone detection in all 21 cases. The level of basal GH varied from 0. 03 μg,/L to 1.50 μg/L. The peak value under GH provocation was from 0. 13 μg/L to 4. 14 μg/L, and the GH was absolute default in all patients. Seventeen cases of them were combined pituitary hormone deficiency (CPHD), and 4 cases were isolated growth hormone deficiency (IGHD). The height of antepituitary was in the range from 1 mm to 3 mm, and the average value was (1.9±1.2) mm. Pituitary stalk was absent in 18 cases and showed as linear and discontinuous stalk in the other 3 cases. The high signal intensity was invisible in normal position in all cases, the high signal intensity spot in the region of infundibular recess of the third ventricle was shown in 19 cases, while it could not be found anywhere in 2 patients with diabetes insipidus. Conclusion PSIS often shows characteristic appearance on MRI, and a definite diagnosis can be made by using MRI combined with clinic hormone detection.
5.Influence of ABCB1 gene polymorphisms on breast cancer with paclitaxel-based chemotherapy
Xinlan LIU ; Haixia ZHANG ; Yueyang ZHAO ; Min JIANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;37(3):383-387
Objective To detect gene polymorphisms of ABCB1/MDR1 in breast cancer and to analyze the association between ABCB1 gene polymorphisms and curative effect of paclitaxel-based chemotherapy in breast cancer.Methods Genotyping of ABCB1exon12 (1236),exon21 (2677)and exon26 (3435)was determined by polymerase chain reaction (PCR)-high resolution melting (HRM)method in 146 cases of female stage IV breast cancer to explore the relationship between the efficacy of chemotherapy in breast cancer patients with paclitaxel chemotherapy.Results In 146 patients with stage IV breast cancer,C1236T CC genotype accounted for 15.86%, CT genotype 44.83%,TT genotype 44.83%,G2677T/A GG genotype 16.67%,GT genotype 45.83%,GA genotype 6.25%,TT genotype 28.47%,AT genotype 2.78%,C3435 CC genotype 22.22%,CT genotype 56.25%,and TT genotype 21.53%.ABCB1 gene polymorphisms did not significantly differ between patients of Hui and Han Nationalities (P>0 .0 5 ).Hardy-Weinberg genetic equilibrium testing showed that polymorphisms of C1236T,G2677T/A and C3435T had group representation (P>0.05).In 146 stage IV breast cancer patients who had received paclitaxel-based chemotherapy,CT/TT genotype of C3435T site had a better response rate (74.11%) (χ2 =12.556,P<0.05),better curative effect than CC genotype (OR=4.183,95% CI 1.838 -9.521,P<0.05),T allele carriers more efficient than C allele carriers with paclitaxel-based chemotherapy (χ2 =8 .5 0 7 ,P<0 .0 5 ). Conclusion Detection of ABCB1 C3435T gene polymorphisms has a significantly clinical value in predicting the efficacy of taxanes chemotherapy in breast cancer patients.
6.Correlation between Polymorphisms of GSTM1, GSTT1 and GSTP1 (rs1695) on Hematologic Toxicities with Anthracycline/Paclitaxel-Based Chemotherapy in Breast Cancer
Xinlan LIU ; Yanjiao ZHAO ; Yin HUANG ; Min JIANG
Tianjin Medical Journal 2014;(8):741-745
Objective To examine the effects of genetic polymorphisms in GSTM1, GSTT1 and GSTP1 (rs1695) genes on hematologic toxicities of breast cancer patients receiving Anthracycline/Paclitaxel- based chemotherapy. Methods Multiply PCR technique and High Resolution Melting method were used to examine these 3 genes polymorphsim in female breast cancers (n=252). Results The GSTP1(rs1695) AG/GG genotype was a risk factor forⅢ-Ⅳdegree of neu-trophil toxicity when patients received Paclitaxel-based chemotherapy and Anthracycline-Paclitaxel-based chemotherapy (OR=6.111, 95%CI 1.526-24.469, P<0.05 and OR=9.257, 95%CI 2.903-29.522, P<0.01). There were no statistic differ-ence onⅢ-Ⅳdegree hematologic toxicities rates between GSTM1(+) and GSTM1(-), GSTT1(+) and GSTT1(-) patients after they receiced Paclitaxel-based chemotherapy and Anthracycline-Paclitaxel-based chemotherapy( P>0.05);There was no statistic difference onⅢ-Ⅳdegree hematologic toxicities rates between GSTM1(+) and GSTM1(-), GSTT1(+) and GSTT1(-), GSTP1AA and GSTP1AG/GG patients after they receiced Anthracycline-based chemotherapy (P>0.05). Conclusion The genetic polymorphisms in GSTP1(rs1695) can be used as a gene marker for forecasting the chemotherapy, inducing neutrope-nia toxicities in breast cancer patients receiving Paclitaxel-based chemotherapy.
7.Effect of TNF-α on production and activation of caspase-3 in primary rat renal proximal tubule cells
Shanying LIU ; Yan LI ; Qiuhui PAN ; Jing WEI ; Xinlan FAN ; Fang SU ; Yanhua LIN ; Tianxin LIN
Chinese Journal of Pathophysiology 2010;26(1):146-149
AIM:To investigate the production and activation of caspase-3 in primary rat renal proximal tubule cells in response to tumor necrosis factor-α(TNF-α) and the implication of nuclear factor-κB (NF-κB) in the process. METHODS:Isolated rat renal proximal tubule cells (PTCs) from male adult Sprague Dawley rats were treated with TNF-α according to the indicated time courses. A specific NF-κB inhibitor,Bay11-7082,was used alone or as a pretreatment for 1 h followed by exposure to TNF-α for 24 h.The protein levels of cleaved caspase-3,caspase-3,I-κBα,phosphorylated I-κBα,and GAPDH were detected by Western blotting using specific antibodies. RESULTS:The protein level of cleaved caspase-3 relative to caspase-3 was significantly increased in the presence of TNF-α for 6 h,12 h,and 24 h. Protein levels of caspase-3 were significantly decreased by 12 h and returned to baseline by 24 h in the presence of TNF-α. Treatment with Bay11-7082 for 25 h alone or pretreatment with Bay11-7082 for 1 h followed by addition of TNF-α for 24 h caused a remarkable reduction in both cleaved caspase-3 and caspase-3 as compared to control and TNF-α treated groups. An increase in phosphorylated I-κBα was observed from 15 min to 60 min after treatment with TNF-α at a dose of 10 μg/L in PTCs. CONCLUSION:NF-κB is not only associated with the activation of caspase-3 but also the production of caspase-3 in primary rat renal proximal tubule cells in response to TNF-α.
8.The effects of saikosaponin-d on the expression of human hepatocellular carcinoma cell BECN1 and autophagic function
Yi XIAO ; Mudan REN ; Guifang LU ; Yan ZHAO ; Dan ZHANG ; Yaping LIU ; Xinlan LU ; Shuixiang HE
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(1):127-130,150
ABSTRACT:Objective To observe the influence of saikosaponin-d (SSd)on the proliferation and the function of autophagy of human hepatocellular carcinoma (HCC)cell line SMMC-7721 to explore the possible mechanisms. Methods SMMC-7721 was cultured invitro and then treated with SSd of various concentrations (5.0,7.5,10.0, 12.5,15.0 and 17.5 mg/L)for 24,48 and 72 h.We used MTT to detect cell proliferation,selected the optimal concentration and time,and detected the expressions of BECN1 at mRNA and protein levels by PCR and Western blot.Results The inhibition rate of the proliferation of SMMC-7721 cell line increased with the increase of the concentration of SSd,and the highest inhibition rate (60%)appeared when the concentration reached 12.5 mg/L. The expression of BECN1 in the group with SSd was obviously higher than that in the control group (P<0.05). 3-MA decreased not only the expressions of BECN1 at mRNA and protein levels but also the expression of BECN1 when used in conjunction with SSd.Conclusion The inhibiting function of SSd on SMMC-7721 presents a dependency between drug concentration and function time,basically in line with the drug dose-effect relationship. SSd induces the occurrence of autophagic cell death through up-regulating the expression of BECN1 ,thus inhibiting the proliferation of SMMC-7 7 2 1 .
9.Association between HBV infection and HLA-DPB1 gene in population of Guangzhou Chinese
Zehuan LIU ; Xinlan FAN ; Jianghai LIN ; Zhiyan FU ; Dejing PAN ; Yonggui FU ; Zongjian JIA ; Anlong XU
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To investigate the association between HBV infection and HLA-DPB1 gene in population of Guangzhou Chinese. METHODS: 58 unrelated patients (test positive of HbsAg,HBeAg,HbcAb) and 75 unrelated healthy control individuals were typed by sequencing based typing (SBT) method in their HLA-DPB1 gene. RESULTS: The phenotype frequencies of HLA-DPB1 alleles of patients and control have no significant difference. CONCLUSION: These results indicate that there is no association between HLA-DPB1 gene and HBV infection.