Comparison of biological behavior of lacrimal gland adenoid cystic carcinoma with high-grade transformation cells

AIM:To evaluate the differences between human lacrimal gland adenoid cystic carcinoma with high-grade transformation(LACC-HGT)primar y cells cultured by high-grade transformation tissue and non-high-grade transformation(non-HGT)primary cells cultured by non-highgrade transformation tissue in proliferation,metastasis,drug susceptibility,and genes.METHODS:LACC-HGT primary cells were established by tissue block culture,and the 4^(th)to 10^(th)generation primary cells were selected as research objects.The cells were preliminarily identified by immunofluorescent staining.The differences between non-HGT and LACC-HGT primary cells in terms of proliferation,metastasis,and drug susceptibility were compared by cell counting kit-8(CCK-8)assay,wound healing,and drug sensitivity experiments.Differentially expressed genes were screened using mRNA array.Gene expression was analyzed using real-time quantitative polymerase chain reaction(RT-qPCR).RESULTS:LACC-HGT primary cells were successfully cultured by tissue block culture.Immunofluorescence staining results showed that cytokeratin(CK)and CK7 expression levels were positive in LACC-HGT primary cells.CCK-8 results showed that the proliferation ability of LACCHGT cells was significantly higher than that of non-HGT cells.Wound healing experiment showed that the migration ability of LACC-HGT cells was significantly higher than that of non-HGT cells.LACC-HGT cells were also less sensitive to cisplatin and paclitaxel than non-HGT cells.Compared with non-HGT cells,9566 differentially expressed genes were found in LACC-HGT primary cells,of which 5162 were upregulated and 4404 were down-regulated.The expression of N-acetylneuraminate pyruvate lyase(NPL),MARVEL domain containing 3(MARVELD3),syntabulin(SYBU),and allograft inflammatory factor 1(AIF1)was higher in LACCHGT cells than in non-HGT cells,whereas that of periostin(POSTN)was lower.CONCLUSION:LACC-HGT primary cells have faster proliferation,stronger migration ability,and poorer sensitivity to chemotherapy drugs than non-HGT primary cells.The expression of mRNAs in non-HGT and LACC-HGT primary cells are significantly different.These features are speculated to be the reasons why high-grade transformation tissues exhibit higher malignant degree and poorer prognosis than their counterparts.

Effects of Xiaoxianxiong Tang on biological behavior in lung cancer cell line A549

Obiective:To investigate the effects of Xiaoxianxiong Tang on the proliferation,invasion,apoptosis,and other cell biological behaviors of non-small cell lung cancer A549 cells.Methods:Human lung adenocarcinoma A549 cells were cultured in vitro,and the IC50 concentration and effective time of administration of Xiaoxianxiong Tang were determined by the CCK-8 assay to detect the inhibitory effect of Xiaoxianxiong Tang on A549 cells proliferation.The effect of the Xiaoxianxiong Tang on apoptosis was determined by flow cytometry;the apoptosis-related protein was detected via Western blot;the metastasis-related protein mRNA was detected by RT-PCR.Results:Xiaoxianxiong Tang significantly inhibited the proliferation viability,the invasive ability,and the clonogenic ability of A549 cells compared with the control group(P<0.001).Moreover,Xiaoxianxiong Tang significantly promoted the apoptosis of A549 cells(P<0.001).Xiaoxianxiong Tang significantly up-regulated Bax and down-regulated Bcl2 expression in A549 cells compared with the control group(P<0.01).The mRNA expression of MMP2 and MMP9 was significantly down-regulated by Xiaoxianxiong Tang compared with the control group(P<0.05).Conclusion:Xiaoxianxiong Tang has the effect of regulating the biological behavior of A549 cells,and Xiaoxianxiong Tang significantly inhibites the proliferation viability,colony formation,and invasion ability of lung cancer A549 cells.

Effects of Curcumin on Biological Behavior and NF-κB/TNF-α Pathway in Mice with Metastatic Bone Pain of Breast Cancer Induced by Walker 256 Cells

Background. The active ingredient curcumin of traditional Chinese medicine was selected as the research object to investigate the possible mechanism of breast cancer metastatic bone pain in mouse walker 256 cells and the effect of curcumin on the NF-κB/TNF-α pathway in order to provide a new idea for clinical treatment of breast cancer metastatic bone pain. Methods. By establishing an animal model of breast cancer bone metastasis in walker 256 cells, the biological behavior of nude mice was observed on the 8th day after successful modeling. Meanwhile, the low dose group, middle dose group and high dose group of mice were given 15 mg/kg, 25 mg/kg, 50 mg/kg of curcumin solution intraperitoneally in 21 days, and the right cavity bone and spinal cord distended in mice (L4-L6) tissues were used to detect related factors, Immunohistochemical method was used to detect c-fos in spinal cord. Expression levels of RANK, NF-κB and TNF-α were detected by RT-PCR and Western blot. Meanwhile, serum levels of Cox2, il-6, leukotriene and PGE2 were detected. Results. Observing the biological behavior index of nude mice, we found that the mechanical pain and thermal pain threshold decreased (p < 0.05), and the cold pain and spontaneous pain scores increased significantly (p < 0.05). After group study, the expression of c-fos in the cancer pain model group was significantly higher than that in the normal control group (p < 0.05), and with the increase of curcumin dose, the expression of c-fos in the high dose group was significantly lower than that in the solvent model group (p < 0.05). The expression of RANK, NF-κB, TNF-α was higher than that of the normal control group and decreased gradually with the increase of curcumin dose, among which the expression of high dose group was significantly lower than that of solvent group (p < 0.05). RANK, NF-κB, TNF-α protein expression was higher than that of normal control group and gradually decreased with the increase of curcumin dose. The levels of Cox2, IL-6, leukotriene and PGE2 in serum decreased with the increase of curcumin dose, and the high dose group decreased significantly (p < 0.05). Conclusions. On the 8th day after the success of the animal model of breast cancer bone metastasis in Walker 256 cells, abnormal biological behaviors such as heat pain, cold pain sensation and spontaneous hyperalgesia were observed. Further studies have found that the increased expression of rank on osteoclasts induced up-regulated expression of NF-κB and c-fos, induced expression of TNF-α gene, and could induce synthesis and release of leukotriene, PGE2 through direct activation of cyclooxygenase, inflammatory media IL-6 cascade reaction, resulting in pathological pain and hypersensitivity. Traditional Chinese medicine active ingredient curcumin could reduce RANK expression of osteoclast, inhibit cell NF-κB and spinal cord c-fos activity, reduce TNF-α expression, inhibit Cox2 activity, and reduce the synthesis and release of inflammatory factors leukotriene and PGE2, thus exerting its analgesic effect, which provides new ideas and methods for clinical treatment of metastatic bone pain in breast cancer.

Changes in biological behaviors of rat dermal f ibroblasts induced by high expression of MMP9

BACKGROUND: The high level of matrix metalloproteinase 9(MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fi broblasts and affect wound healing is still unclear. The present study aimed to observe changes in the biological behaviors of rat dermal fi broblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot.METHODS: A cell model of skin f ibroblast with high expression of MMP9 was established by coculture of high glucose(22.0 mmol/L) and homocysteine(100 μmol/L). A control group was incubated with normal glucose(5.5 mmol/L). Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwell were used to detect cell proliferation, viability, collagen(hydroxyproline) secretion, horizontal migration and vertical migration of cells. The data were expressed as mean±SD. P value less than 0.05 was considered statistically signif icant.RESULTS: The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group(7.05±1.02 vs. 1.00±0.00, 206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively, P<0.01). The proportion of S-phase cells, proliferation index, cell viability, collagen(hydroxyproline) secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group(P<0.01).CONCLUSION: Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fi broblasts.

Effects of targeted-edited oncogenic insulin-like growth factor-1 receptor with specific-sgRNA on biological behaviors of Hep G2 cells

BACKGROUND Insulin-like growth factor-1 receptor(IGF-1R)is over-expressed in hepatocellular carcinoma(HCC).However,the relationship between IGF-1R activation and HCC progression remains unidentified.AIM To investigate the effects of editing IGF-1R on the biological features of HCC cells.METHODS Immunohistochemistry analyzed the expressions of IGF-1R and P-glyco protein(P-gp)in HCC tissues and their distal non-cancerous tissues(non-Ca).IGF-1R was edited with Crispr/Cas9 system,screened specific sg RNAs,and then transfected into Hep G2 cells.CCK-8,scratch wound test detected cell proliferation,migration,invasion and transwell assays,respectively.Alterations of IGF-1R and P-gp were confirmed by Western blotting.Alterations of anti-cancer drug IC_(50)values were analyzed at the cell level.RESULTS The positive rates of IGF-1R(93.6%,χ~2=63.947)or P-gp(88.2%,χ~2=58.448)were significantly higher(P<0.001)in the HCC group than those(36.6%in IGF-1R or 26.9%in P-gp)in the non-Ca group.They were positively correlated between high IGF-1R and P-gp expression,and they were associated with hepatitis B virus infection and vascular invasion of HCC.Abnormal expressions of circulating IGF-1R and P-gp were confirmed and associated with HCC progression.Biological feature alterations of HCC cells transfected with specific sg RNA showed IGF-1R expression down-regulation,cell proliferation inhibition,cell invasion or migration potential decreasing,and enhancing susceptibility of Hep G2 cells to anti-cancer drugs.CONCLUSION Edited oncogenic IGF-1R was useful to inhibit biological behaviors of Hep G2 cells.

Effect of lentiviral expression of circLIFR on the biological behavior of Hep3B hepatocellular carcinoma cells

Objective:To investigate the effect of lentiviral stable high expression of circLIFR on the biological behavior of Hep3B hepatocellular carcinoma(HCC)cells.Methods:Hep3B cell lines were infected with lentiviral packaging of circLIFR expression plasmids to construct stable circLIFR high expression HCC cells.The lentivirus infected with circLIFR high expression sequence was used as the circLIFR high expression group,the lentivirus infected with circLIFR high expression empty vector sequence was used as the negative control group,and the uninfected group was used as the blank control group.After that,circLIFR expression levels were detected by qPCR,and the back splice sites were identified by Sanger sequencing.The cell viability was examined by cell proliferation kit and invasive ability was determined by Transwell assay.Results:The qPCR and Sanger sequencing showed that the stable circLIFR expression of Hep3B cells was successfully established.The circLIFR high expression group had better cell proliferation viability than the negative control and blank control groups,and the differences were statistically significant(P<0.05).The number of cells crossing Matrigel gel in the negative control and blank control groups was(270.8±18.9)and(266.2±17.6),respectively,while the number of cells crossing Matrigel gel in the circLIFR high expression group was(396.6±32.9),and the differences were statistically significant(P<0.05).Conclusion:High circLIFR expression considerably promotes the proliferation and invasive ability of Hep3B cells.

Biological behaviors of two novel syngeneic human osteosarcoma cell lines

Objective To characterize and compare the different biological behaviors of two novel human osteosarcoma cell lines,Zos and Zos-M,established respectively from the primary site and the skip metastasis of an osteosarcoma patient.Methods Two

MiR-183-5p promotes the progression of non-small cell lung cancer through targeted regulation of FOXO1

Objective To investigate miR-183-5p targeting to forkhead box protein O1(FOXO1)and its corresponding effect on the proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT)of non-small cell lung cancer(NSCLC)cells.Methods NSCLC tissues and adjacent normal tissues from 60 patients with NSCLC adenocarcinoma were obtained via pathological biopsy or intraoperative resection.Several cell lines were cultured in vitro,including the human normal lung epithelial cell line BEAS-2B and human NSCLC cell lines A549,SPCA-1,PC-9,and 95-D.miR-183-5p and FOXO1 mRNA expression in tissues and cells were detected by qRT-PCR;the corresponding correlations in NSCLC tissues were analyzed using the Pearson test,and the relationship between miR-183-5p expression and clinicopathological parameters was analyzed.The miR-183-5p-mediated regulation of FOXO1 was verified by bioinformatics prediction alongside double luciferase,RNA-binding protein immunoprecipitation(RIP)assay,and pull-down experiments.A549 cells were divided into control,anti-miR-NC,anti-miR-183-5p,miR-NC,miR-183-5p,miR-183-5p+pcDNA3.1,and miR-183-5p+pcDNA3.1-FOXO1 groups.Cell proliferation,invasion,migration,apoptosis,and cell cycle distribution were detected using an MTT assay,clone formation assay,Transwell assay,scratch test,and flow cytometry,respectively.The expression of EMT-related proteins in the cells was analyzed by western blotting.The effect of miR-185-3p silencing on the development of transplanted tumors was detected by analyzing tumor formation in nude mice.Results miR-183-5p expression was significantly higher in NSCLC tissues and cells than in adjacent normal tissues,whereas FOXO1 mRNA expression was significantly down-regulated.There was a significant negative correlation between miR-183-5p and FOXO1 mRNA in NSCLC tissues(P<0.05).Additionally,the expression of miR-183-5p was significantly correlated with tumor size,tumor differentiation,and tumor-node-metastasis stage in patients with NSCLC(P<0.05).miR-183-5p targeted and inhibited FOXO1 expression.Compared to the anti-miR-NC group,the cell proliferation,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells were significantly lower in the anti-miR-183-5p group,whereas the protein expression of E-cadherin andα-catenin and the proportion of G0/G1 phase cells were significantly higher;additionally,the frequency of colony formation and invasion were significantly lower in the anti-miR-183-5p group(P<0.05).Compared to the miR-NC group,the cell proliferation,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells in the miR-183-5p group were significantly higher,whereas the E-cadherin andα-catenin protein expression and the proportion of G0/G1 phase cells were significantly lower;furthermore,the frequency of colony formation and invasion were significantly higher in the miR-183-5p group(P<0.05).Compared with the miR-183-5p+pcDNA3.1 group,the OD value,scratch healing rate,N-cadherin and vimentin protein expression,and the proportion of S phase cells were significantly lower in the miR-183-5p+pcDNA3.1-FOXO1 group,whereas E-cadherin andα-catenin protein expression and the proportion of G0/G1 phase cells were significantly higher;additionally,the frequency of colony formation and invasion was significantly lower in the miR-183-5p+pcDNA3.1-FOXO1 group(P<0.05).Overall,silencing miR-185-3p inhibited the growth of transplanted tumors and promoted FOXO1 expression.Conclusion Overexpression of miR-183-5p can inhibit apoptosis and promote the proliferation,migration,invasion,and EMT,of NSCLC cells by down-regulating FOXO1 expression.

Impact of COVID-19 pandemic on clinicopathological features of transplant recipients with hepatocellular carcinoma:A case-control study

BACKGROUND The coronavirus disease 2019(COVID-19)pandemic had a significant impact on the management of all diseases.Various diseases such as cancer have a higher risk of COVID-19-related death.Despite this fact,any delay or alteration in treatment of cancer may have fatal consequences.Hepatocellular carcinoma(HCC)is an aggressive liver cancer that requires multimodality treatment to improve survival.AIM To evaluate the impact of COVID-19 on the management of patients with HCC by determining changes in demographic,clinical and histopathological variables.METHODS Demographic,clinical and pathological variables of patients with HCC who had undergone liver transplantation between March 2020 and June 2021(Pandemic group,n=48)were retrospectively compared with that of the patients with HCC transplanted between November 2018 and March 2020(Pre-pandemic group,n=61).RESULTS The median age of the patients in the study was 56(interquartile range=15).Ninety-seven patients(89%)were male and 12 were female(11%).The most common etiology of liver disease was hepatitis B virus(n=52,47.7%).According to our results,there was a 21.3% drop in the number of patients transplanted for HCC.There was no difference in the demographic,clinical and pathological characteristics of the patients except blood alkaline phosphatase levels(P=0.029),lymphovascular invasion(P=0.019)and type of the liver graft that was transplanted(P=0.017).CONCLUSION It is important to develop a surveillance strategy for liver transplant centers.The liver transplantation for HCC is justified and safe provided that strict surveillance protocols are applied.

Adverse effects of chemoradiotherapy on invasion and metastasis of tumor cells

The phenomenon of enhanced invasion and metastasis of residual tumor cells has been observed in an increasing number of patients receiving chemoradiotherapy recently,and tumor metastasis will undoubtedly limit patient prognosis.However,the key mechanism by which chemoradiotherapy affects the invasion and metastasis of tumor cells remains unclear.Studies have shown that chemoradiotherapy may directly act on tumor cells and alter the tumor microenvironment,or induce cell apoptosis and autophagy to promote tumor cell survival and metastasis.In this review,we summarize the potential mechanisms by which chemoradiotherapy may affect the biological behavior of tumor cells and open up new avenues for reducing tumor recurrence and metastasis after treatment.These insights will improve the efficacy of chemoradiotherapy.

Gastrodin modified polyurethane conduit promotes nerve repair via optimizing Schwann cells function

Peripheral nerve regeneration and functional recovery remain a major clinical challenge.Nerve guidance conduit(NGC)that can regulate biological behavior of Schwann cells(SCs)and facilitate axonal regeneration through microenvironmental remodeling is beneficial for nerve regeneration and functional recovery.Gastrodin,a main constituent of a Chinese traditional herbal medicine,has been known to display several biological and pharmacological properties,especially antioxidative,anti-inflammatory and nerve regeneration.Herein,polyurethane(PU)NGCs modified by different weight ratio of Gastrodin(0,1 and 5 wt%)were designed for sequential and sustainable drug release,that created a favorable microenvironment for nerve regeneration.The scaffold showed suitable pore structure and biocompatibility in vitro,and evidently promoted morphological and functional recovery of regenerated sciatic nerves in vivo.Compared to the PU and 1% Gastrodin/PU scaffolds,the 5% Gastrodin/PU significantly enhanced the proliferation,migration and myelination of SCs and up-regulated expression of neurotrophic factors,as well as induction of the differentiation of PC12 cells.Interestingly,the obvious anti-inflammatory response was observed in 5% Gastrodin/PU by reduced expression of TNF-α and iNOS,which also evidenced by the few fibrous capsule formation in the subcutaneous implantation.Such a construct presented a similarity to autograft in vivo repairing a 10 mm sciatic nerve defects.It was able to not only boost the regenerated area of nerve and microvascular network,but also facilitate functional axons growth and remyelination,leading to highly improved functional restoration.These findings demonstrate that the 5% Gastrodin/PU NGC efficiently promotes nerve regeneration,indicating their potential for use in peripheral nerve regeneration applications.

Imaging biomarkers for predicting poor prognosis of hepatocellular carcinoma: a review

Hepatocellular carcinoma(HCC)is a primary malignancy of the liver with a high mortality rate.Heterogeneity is the main biological characteristic of HCC,which manifests through the different biological behaviors of each phenotype and ultimately,affects patient prognosis and treatment efficacy.Various aggressive biological behaviors are considered to be associated with the poor prognosis of HCC patients including poor differentiation,microvascular invasion,intracellular fat accumulation,invasive growth,bile duct invasion or tumor thrombosis,and tumor spread and metastasis,and have been reported as prognostic biomarkers.In addition,HCC results from multifactor synergistic damage,and various factors related to genetics,molecular pathology and immunohistochemistry such asβ-catenin,Ki67,cytokeratin-19,and epithelial cell adhesion molecule have an impact on HCC differentiation and prognosis.This article is an overview of the biological behaviors that lead to poor prognosis of HCC,and the roles of morphological and quantitative noninvasive imaging biomarkers in the evaluation and prediction of these behaviors.Some common biomarkers related to genetics,molecular pathology and immunohistochemistry are also briefly summarized.It is hoped that this review will provide clinicians and radiologists with an update on the development of liver imaging,and provide directions for the combination of radiology,genetics,molecular pathology and histopathology to better predict the prognosis of HCC patients.