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Expression of Notch signaling pathway in pancreatic cancer and its role

Gan LUO ; Chao YI ; Abula YIMAMUMAIMAITIJIANGO ; Lin XU ; Jiwei YIN ; Wei DING

Practical Oncology Journal.2019;33(1):1-7. doi:10.11904/j.issn.1002-3070.2019.01.001

Objective The objective of this study was to investigate the abnormal expression of Notch signaling pathway members in pancreatic cancer and its important effect on the development of pancreatic carcinoma. Methods Affymetrix gene expres-sion microarray was used to screen the differentially expressed members of Notch signal pathway in 10 cases of pancreatic carcinoma and its adjacent tissues,and verified by real-time quantitative PCR and Western blotting. The lentivirus expression vector carrying the siRNA fragment of Jagged 2(JAG2)gene was transfected into the pancreatic cancer primary cells to construct the JAG2 gene re-pression-expressing pancreatic cancer cell line. MTT,flow cytometry and Transwell assays were used to analyze cell proliferation, changes of cell cycle and invasive transfer capabilities. Results A total of 512 differentially expressed genes were detected by Affy-metrix gene expression microarray,including 419 up-regulated genes and 93 down-regulated genes. JAG2( up-regulated expres-sion 8. 20 times),NOTCH1(up-regulated expression 3. 74 times),HES1(up-regulated expression 3. 27 times),and NOTCH2(up-regulated expression 3. 16 times)were differentially expressed in Notch signaling pathway. The results of PCR and Western blotting were consistent with those of gene chip. The growth curves of JAG2 gene repressed pancreatic cancer cells and pancreatic cancer pri-mary cells were drawn by the standard OD490 value of d1-d5 by MTT assay. JAG2 gene repressor expression vector could signifi-cantly inhibit the proliferation of pancreatic cancer cells. The cell cycle analysis showed that the apoptosis and the arrested cell cycle at the S phase were significantly increased in pancreatic cancer cells with JAG2 gene repressor expression. The invasive ability was significantly reduced in JAG2 gene repressor expression pancreatic cancer cells(P<0. 05). Conclusion Some members of the Notch signaling pathway are significantly differentially expressed in pancreatic cancer tissues,and repression of this member can affect the growth,cell cycle,invasion and metastasis of pancreatic cancer cells.

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Research on methylation of FHIT,hMLH1,p16,RAR-beta,Reprimo and TIMP3 genes in gastric cancer patients

Yuxian BAI ; Zhongjuan MA ; Yingling SU ; Anguo MU ; Rui XIE

Practical Oncology Journal.2019;33(1):8-13. doi:10.11904/j.issn.1002-3070.2019.01.002

Objective The aim of this study was to investigate the methylation status of fragile histidine triad( FHIT) gene, human mutl homolog 1(hMLH1)gene,p16 gene,retinoic acid receptor beta( RAR-beta) gene,Reprimo gene and tissue inhibitor of metalloproteinase 3 ( Timp3) gene in gastric cancer and corresponding paracancerous tissues. Methods The methylation levels of FHIT,hMLH1,p16,RAR-beta,Reprimo and TIMP3 genes in 42 clinically resected gastric cancer specimens and 42 corresponding paracancerous tissues were detected by sodium bisulfite sequencing. Results The average methylation rates of the genes in gastric cancer and corresponding paracancerous tissues were:FHIT(1. 50% ,1. 36% ),hMLH1(4. 77% ,0. 48% ),p16(9. 63% ,10. 36% ), RAR-beta(4. 75% ,4. 17% ),Reprimo(9. 71% ,3. 76% )and TIMP3 genes(18. 34% ,14. 06% ). Compared with the paracancerous control group,the average methylation rate of Reprimo gene was only statistically different in gastric cancer patients(P=0. 00787). The difference in methylation rate of Reprimo gene promoter in gastric cancer patients with the degree of tissue differentiation was sta-tistically significant(P<0. 05). Conclusion There has methylation in the cytosine guanidine dinucleotide island of the Reprimo gene promoter region in gastric cancer. The high methylation rate of the Reprimo gene can be used as a potential biomarker for gastric cancer to detect the early stage of gastric cancer.

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Mechanism of tumor suppressor gene DKK2 inhibiting proliferation of nephroblastoma cells in children

Guixia YUAN ; Yan LI ; Wanqi FENG ; Xiaojuan XIA ; Xu LI

Practical Oncology Journal.2019;33(1):14-20. doi:10.11904/j.issn.1002-3070.2019.01.003

Objective The aims of this study were to investigate the expression of DKK2,a WNT signaling pathway regula-tor,in nephroblastoma cells and tissues of children,the effect on the proliferation of nephroblastoma SK-NEP-1 cells,and to explore its mechanism. Methods The relative expression of DKK2 in nephroblastoma cells and tissues was analyzed by qRT-PCR and im-munoblotting assays. Overexpressing DKK2 SK-NEP-1 cells were set as the experimental group( DKK2 group);the blank control plasmid group was set as a control group( Vector group),transfected with pcDNA3. 1 ( +) -Flag-DKK2 plasmid( Experimental group)and pcDNA3. 1( +) -Flag-Vector plasmid(Control group). The over-expression of DKK2 was confirmed in SK-NEP-1 cells by RT-PCR and immunoblotting. CCK-8 and cell cloning assays were used to determine the effect of DKK2 on cell prolifera-tion;flow cell cycle and apoptosis assays were used to confirm the effect on cell proliferation in overexpressed DKK2 cells. The xen-graft formation assay in nude mice was to verify the effect of DKK2 on proliferation in overexpressed DKK2 cells;the mechanism of DKK2 in inhibitory proliferation was analyzed by qRT-PCR,Western blotting and immunohistochemistry. Results Compared with normal renal epithelial tissues,DKK2 mRNA was down-regulated in children with nephroblastoma,and the difference was statistically significant(P<0. 001). Compared with the control group,transfected DKK2 cell viability was significantly inhibited after treatment for 24,48 and 72 h( P<0. 05),cell clone formation in the experimental group was significantly inhibited(31. 11% ± 2. 14% ) ( P<0. 05),the cell cycle in the experimental group was significantly arrested at the G1 phase(P<0. 001),and the apoptosis rate in the experimental group was significantly increased(P<0. 001). Compared with the control group,the tumor weight and volume in nude mice were significantly low in the experimental mice which were injected DKK2 overexpression cells(P<0. 001). Active-β-cate-nin and downstream genes were significantly inhibited in over-expressed DKK2 SK-NEP-1 cells. Conclusion DKK2 is down-regulated in human cutaneous nephroblastoma and participates in the mechanism of nephroblastoma by antagonizing Wnt/β-catenin signaling pathway.

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Effect of miR-129 on proliferation and apoptosis of osteosarcoma MG-63 cells by targeting SEPHS1

Chong CHEN ; Jian WANG ; Shuai YANG

Practical Oncology Journal.2019;33(1):21-26. doi:10.11904/j.issn.1002-3070.2019.01.004

Objective The aim of this study was to investigate the effect of miR-129 on the proliferative activity and apop-tosis of osteosarcoma MG-63 cells. Methods Thirty cases of osteosarcoma and its adjacent paracancerous tissues were collected. RT-PCR was used to detect the expressions of miR-129 mRNA and SEPHS1 mRNA. Western blot was used to detect the protein level of SEPHS1. After MiR-129 was over-expressed and knocked down,the cell proliferation was detected in MG-63 cells by CCK-8,and apoptosis was detected in MG-63 cells by Westerm blot and hoechest staining. Results In this study,the expression of miR-129 in osteosarcoma tissues was significantly higher than that in para-cancerous tissues(P<0. 05). Through the established model in vitro of miR-129 overexpression or knockdown,it was determined that the miR-129 mimetic and inhibitor concentrations were optimal for overexpression and knockdown at 50 nM and 200 nM, respectively. The possibility of binding between miR -129 and SEPHS1 was predicted by software,and the luciferase reporter assay further confirmed the binding relationship between miR-129 and SEPHS1. Knockdown of miR-129 significantly inhibited the proliferation of MG-63 cells and accelerated the apoptosis of MG-63 cells. Conclusion miR -129 plays a key role in regulating the proliferation and apoptosis of osteosarcoma cells by binding to SEPHS1.

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Changes of serum CRP and hs-CRP levels in patients with liver cancer and depression and their influence on prognosis

Fengjuan ZHAO ; Qi WANG ; Yunhong REN ; Jueling WEI ; Lequn LI ; Xinhua ZHAO ; Juan TANG ; Xuemei YOU

Practical Oncology Journal.2019;33(1):27-33. doi:10.11904/j.issn.1002-3070.2019.01.005

Objective The objective of this study was to investigate the effect of depression on serum levels of C-reactive protein(CRP)and high-sensitivity C-reactive protein(hs-CRP),and prognosis in liver cancer patients. Methods A total of 251 patients with liver cancer undergoing hepatectomy were enrolled. The hospital anxiety and depression scale( HADS-D) and 9-item patients health questionnaire(PHQ9) were assessed for depression before 3 days for surgery. Patients were divided into depression group(n=95)and non-depression group(n=156) according to the scores. Preoperative serum levels of CRP,hs-CRP,ALT and AST were measured and compared between the depression and non-depression groups. Survival analysis Kaplan-Meier method was used to compare the disease-free survival(DFS)and total survival(OS)between the two groups. Results The serum levels of CRP, hs-CRP,ALT and AST in the depression group were significantly higher than those in the non-depression group(P<0. 05). The follow-up of 3. 5-year showed that 164 patients(65 in depression group and 99 in non-depression group)had recurrence or metas-tasis and 47 patients(22 in depression group and 25 in non-depression group) died. The DFS and OS in the depression group were significantly lower than those in the non-depression group(P< 0. 05). Cox multiple regression analysis showed that liver function grading,BCLC staging and depression were independent risk factors for the prognosis of liver cancer. Spearman correlation analysis showed that patients′degree of depression was positively correlated with serum levels of CRP and hs-CRP(P<0. 05),DFS and OS were negatively correlated with serum levels of CRP and hs-CRP(P<0. 05). Conclusion Depression may mediate elevated serum levels of CRP and hs-CRP,maintain inflammatory response in patients,lead to increased liver function damage,elevate levels of ALT and AST,and thus adversely affect the prognosis of patients with liver cancer.

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Prognostic significance of neutrophil/lymphocyte ratio and platelet/lymphocyte ratio in systemic inflammation-related indi-cators in gastric cancer patients after radical resection

Fengchun ZHANG ; Ying JIANG ; Zhaonan LIU ; Ningning YAN ; Hongquan CUI ; Yingchun XU

Practical Oncology Journal.2019;33(1):34-39. doi:10.11904/j.issn.1002-3070.2019.01.006

Objective The aim of this study was to compare the short-term and long-term prognostic significance of dif-ferent systemic inflammatory scores in patients with gastric cancer:neutrophil lymphocyte ratio(NLR) and platelet lymphocyte ratio ( PLR). Methods The clinical data of 240 patients with gastric cancer who underwent radical surgery were retrospectively analyzed. The relationship between NLR,PLR,glasgow prognostic score( GPS) and clinicopathological characteristics and perioperative compli-cations were compared. Survival analysis was performed using Kaplan-Meier survival analysis. The Log-rank methods were used to test the difference significance,and the multivariate analysis was performed using the Cox regression risk model. Results Patients in the high NLR and PLR groups were older,had a high GPS,deep tumor infiltration,more lymph node metastasis,and a late TNM stage (P<0. 05). The rate of positive margin in the high NLR group was higher(P<0. 05). Patients in the high NLR and PLR groups re-quired significantly higher transfusion rates than those in the low NLR and PLR groups(P<0. 05),and the high NLR group had more postoperative complications(P<0. 05). Univariate analysis showed that age,GPS,tumor location,tumor infiltration depth,lymph node metastasis status,TNM stage,vascular tumor thrombus,nerve infiltration,NLR and PLR were associated with postoperative survival of gastric cancer patients(P<0. 05). The overall survival(OS)in high NLR and high PLR groups was lower than that of low NLR and low PLR groups(P=0. 018 and P<0. 001). Cox regression analysis showed GPS and lymph node metastasis were independent prog-nostic factors of OS(P<0. 001 and P=0. 002). Conclusion Preoperative systemic inflammatory scores NLR and PLR are prognos-tic factors affecting the preoperative clinical outcomes of gastric cancer. It is recommended to be used in combination with other prog-nostic indicators for routine use in the prognosis of patients undergoing radical gastrectomy.

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Clinical significance of combined detection of PD-L1 protein and CD8 +TILs in resected lung adenocarcinoma

Jianli MA ; Minghui ZHANG ; Hongfei JI ; Xiangying XU

Practical Oncology Journal.2019;33(1):40-46. doi:10.11904/j.issn.1002-3070.2019.01.007

Objective The prognostic role of PD-L1 expression in surgically resected lung adenocarcinoma(ADC)remains controversial. The aim of present study was to investigate the prognostic value of PD-L1 combined with CD8 +TILs expression in pa-tients with resectable lung adenocarcinoma. Methods A total of 104 patients with lung adenocarcinoma who underwent radical resec-tion were enrolled at the Affiliated Tumor Hospital of Harbin Medical University from January 2009 to December 2012. Immunohisto-chemistry was used to detect the expression of PD-L1 and CD8 +TILs in primary tumor specimens. Its clinical pathological factors and its relationship with prognosis were statistical analysis. Results The positive expression rate of PD -L1 in tumor cells was 38. 5% (40/104),and the positive rate of CD8 +TILs was 44. 2% (46/104). There was a significant correlation between the expres-sion of CD8 +TILs and TNM stages of patients with resectable ADC. The Kaplan-Meier survival analysis showed that low expression of PD-L1 and high expression of CD8 +TILs were significantly associated with disease -free survival ( DFS) and overall survival ( OS),and could be used as an independent predictors for predicting prognosis of patients with resectable ADC. Conclusion The ex-pression of PD-L1 and CD8 +TILs in the postoperative specimens can help to judge the prognosis of patients. This study has certain significance for immunotherapy of PD-1/PD-L1 pathway in patients with surgically resected ADC.

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Correlation between HER-2 receptor of progesterone and estrogen receptor-positive breast cancer and mammography imaging & clinicopathological features

Zhingying MA ; Naijian SHANG ; Miaotian CAI ; Yang SHEN ; Zhiyun JIANG

Practical Oncology Journal.2019;33(1):47-51. doi:10.11904/j.issn.1002-3070.2019.01.008

Objective The objective of this study was to investigate the relationship between the molybdenum target imaging features and clinical signs of mammography,and the expression of HER-2 receptor in breast cancer patients with positive receptors of estrogen and progesterone. Methods One hundred and eleven patients with estrogen and progesterone receptor-positive breast canc-er who were admitted to our hospital from July to September 2017 were examined with mammography before operation. Immunohisto-chemical analysis was performed on breast cancer tissues. Pathological features and features related to mammography were analyzed af-ter operation. The relationship between clinicopathological features and mammography-related signs,and HER-2 expression was al-so analyzed statistically. Results Of the 111 patients,34 were HER-2 positive patients. There were significant difference between the observed indicators and the over-expression of HER-2(P<0. 05). These indicators included lymph node metastasis,calcifica-tion of the mass,margin of the mass,the shape of calcification,and the blood status of vessels. The mammography of patients with tri-ple positive breast cancer is more likely to manifest as lymph node enlargement,mass with calcification or simple calcification,margin-al burrs,granular calcification,and vascular thickening. Conclusion The expression of HER-2 in breast cancer patients with posi-tive estrogen and progesterone receptors can be reflected to some extent by lymph node metastasis,tumor with calcification,tumor mar-gin,calcification,calcification morphology and peripheral blood vessel thickening. In terms of clinical conditions,The biological behav-ior and characteristics of tumor can be inferred based on the clinicopathological features of the patient and the imaging findings of the mammography,providing a direction for the treatment and evaluation of prognosis in breast cancer patients.

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Expression of COL5A2 in bladder cancer tissues and its correlation with clinicopathological features and prognosis

Hongbo YANG ; Jie SHI ; Yong YE ; Wencheng LI ; Zhaoping DENG

Practical Oncology Journal.2019;33(1):52-56. doi:10.11904/j.issn.1002-3070.2019.01.009

Objective The aim of this study was to investigate the expression of COL5A2 in bladder cancer tissues,and its correlation with clinicopathological features and prognosis. Methods A total of 144 patients with bladder cancer were enrolled in this study. Real-time fluorescence reverse transcription and immunohistochemistry were used to detect the expression of COL5A2 at levels of mRNA and protein in bladder cancer tissues and normal bladder tissues. The relationship between COL5A2 expression and clinico-pathological features and prognosis was analyzed. Results The expression of COL5A2 mRNA in the bladder cancer group was higher than that in the paracancerous group(P<0. 05). The positive rate of COL5A2 in the bladder cancer group was higher than that in the normal bladder tissues(P<0. 05). The expression of COL5A2 protein was not correlated with age( P>0. 05),positively associated with the TMN stage,pathological grade,tumor maximum diameter(≥5 cm),depth of invasion,lymph node metastasis,lymphatic vas-cular infiltration,and recurrence;the difference was statistically significant(P<0. 05). The 3-year survival rate and survival time in the COL5A2 negative group were significantly higher than those in the COL5A2 positive group(P<0. 05). The higher TMN stage,the higher pathological stage,the maximum diameter of the tumor(≥5 cm),the deeper infiltration depth,lymph node metastasis,lymphat-ic vascular infiltration,recurrence,the higher positive expression rate of COL5A2 protein. Conclusion COL5A2 is highly expressed in the bladder cancer tissues, which promotes the development of bladder cancer. Bladder cancer patients with low expression of COL5A2 can obtain a good prognosis.

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Therapeutic effect of minimally invasive small incision surgery on patients with early thoracic esophageal cancer and its in-fluence in pain

Zhongyang LIU ; Xin YUAN ; Wen ZHOU ; Xiaoliang LI ; Limao FAN

Practical Oncology Journal.2019;33(1):57-61. doi:10.11904/j.issn.1002-3070.2019.01.010

Objective The aim of this study was to explore the effect of minimally invasive small incision surgery on early thoracic esophageal cancer patients and its impact on pain. Methods A total of 160 patients with early thoracic esophageal cancer who were treated in our hospital from March 2016 to March 2017 were randomly divided into the conventional surgery group and the minimally invasive small incision surgery group. Patients in the conventional surgery group were treated with routine operation,and pa-tients in the minimally invasive small incision surgery group were treated with minimally invasive small incision surgery. Vital capacity ( VC),forced expiratory volume 1(FEV1),visual analogue scale(VAS),hospitalization time,operation time and intraoperative bleed-ing volume were counted. Growth hormone(GH)was detected by ELISA. The levels of cortisol(Cor) and interleukin-8( IL-8) were measured by immunoassay and adverse reactions. Results The levels of VC and FEV1 in the minimally invasive small incision group were significantly higher than those in the conventional surgery group. The bleeding volume,operation time and hospitalization time were shorter than those in the conventional surgery group. The levels of GH and Cor,VAS score and incidence of adverse reaction rate were lower than those in the conventional surgery group,and the level of IL-8 was also higher in than that in the conventional surgery group. The difference was statistically significant(P<0. 05). Conclusion Minimally invasive small incision surgery is effective on the treatment of early thoracic esophageal cancer,relieving pain and reducing adverse reactions.

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