Influence of recombinant retroviral vector expressing antisense TGF alpha on malignant phenotype of human pancreatic carcinoma cell line.

OBJECTIVE To observe the effects of antisense TGF alpha on the growth of human pancreatic carcinoma cell line cells.

METHODS A recombinant retroviral vector expressing antisense TGF alpha was constructed, and transfected the ecotropic packaging cell line psi-2 with lipofectin. After the amphotropic packaging cell line PA317 was transfected with the virus supernatant of psi-2, the replication-defective, amphotropic retroviral supernatant was used to infect human pancreatic carcinoma cell line PC-7. Following puromycin selection, puromycin-resistant colonies were pooled and expanded to a cell line PC-7/AS-TGF alpha.

RESULTS The retroviral integration in the genomes of psi-2, PA317 and transformant PC-7 cells was confirmed by Southern blot hybridization. Northern blot hybridization showed a down regulation of endogenous TGF alpha in PC-7/AS-TGF alpha cell line. The high levels of growth inhibition and reduction of 3H-TdR incorporation in PC-7/AS-TGF alpha were evident. Also, the soft agar colony-formation and tumorigenicity in nude mice were significantly suppressed by antisense TGF alpha.

CONCLUSIONS The antisense TGF alpha expressing vector can block the target gene expression, suppress the cell growth and partially reverse the malignant phenotype of pancreatic carcinoma cells.

     

Effects of antisense epidermal growth factor and its receptor retroviral expression vectors on cell growth of human pancreatic carcinoma cell line.

A 150 bp epidermal growth factor (EGF) cDNA fragment and a 1024 bp epidermal growth factor receptor (EGFR) cDNA fragment were inserted into 5.05 kb pBabe-puro retroviral vectors between BamH I and EcoR I sites in 3'-5' and / or 5'-3' orientation. The vectors were ligated with EGF and EGFR fragments by T-4 Ligase. The recombinant retroviral vectors were then packaged with packaging cell line PA317 through calcium phosphate mediated transfection. The viral supernatant of transfected PA317 cell lines were used to infect the human pancreatic carcinoma cell line PC-7. The resultant transformant cell lines: PC-7 / AS-EGF, PC-7 / S-EGFR, PC-7 / AS-EGFR and PC-7 / pBabe were tested for their endogenous EGF and EGFR mRNA expressions, cell growth rate, 3H-TdR incorporation rate, soft agar colony formation and tumorigenicity in nude mice. The results showed that there were noticeable inhibitions of cell growth, 3H-TdR incorporation rate, soft agar colony formation and tumorigenicity in nude mice in PC-7 / AS-EGF an     

Effects of apigenin on cell proliferation of human pancreatic carcinoma cell line BxPC-3 in vitro

Objective： To observe the effects of apigenin on cell proliferation of human pancreatic carcinoma cell line BxPC-3 in vitro. Methods：The inhibitive effects of apigenin at different concentrations （0 μmol/L, 100 μmol/L, 200 μmoL/L, and 400 μmol/L） on human pancreatic carcinoma cell line BxPC-3 were detected by MTT assays, transmission electron microscope, agarose gel electrophoresis and flow cytometry. The immunohistochemistry was used to detect the expression of Bcl-2 and Bax gene. Results： Apigenin at different concentrations could inhibit the proliferation of human pancreatic carcinoma cell lines BxPC-3, and the inhibitive effect was dose-dependent. The cell cycle of pancreatic carcinoma cells was arrested at G2/M phase. The results of immunohistochemistry showed that the density of apigenin increased, and the expression of Bcl-2 gene was reduced gradually, At the same time the expression of Bax gene was enhanced. Conclusion ：Apigenin could inhibit the proliferation of human pancreatic carcinoma cell lines BxPC-3 in vitro. The effect of apoptosis was accompanied with the expression of Bcl-2 decrease and Bax increase.     

Modulating effects of survivin antisense oligonucleotide on changes of apoptosis and cell cycle of human hepatocellular carcinoma cell line SMMC-7721

Modulating effects of survivin antisense oligonucleotide on changes of apoptosis and cell cycle of human hepatocellular carcinoma cell line SMMC-7721@陈涛$Gener Surg Center,Gener Hosp Chengdu Command,Chengdu 610083     

Inhibiting effect of antisense hTRT on telomerase activity of human liver cancer cell line SMMC-7721

Objective: To induce changes in biological character of human liver cancer cell line SMMC-7721 by blocking the expression of telomerase genes hTRT and to explore its value in cancer gene therapy. Methods: The vehicle for eukaryotic expression of antisense hTRT was constructed and then transfected into SMMC-7721 cells. The effects of antisense hTRT gene on telomerase activity, cancer cell growth and malignant phenotypes were analyzed. Results: The obtained transfectants that could express antisense hTRT gene stably showed marked decrease in telomerase activity;the shortening of telomere was obvious; cells presented contact growth inhibition; in nude mice transplantation, the rate of tumor induction dramatically decreased. Conclusion : Antisense hTRT gene expression can significantly inhibit telomerase activity of cancer cells and decrease malignant phenotypes in vitro and in vivo. Therefore, as a telomerase inhibitor, antisense hTRT gene may be a new pathway for cancer therapy.     

Killing effect of adenoviral mediated cytosine deaminase gene on human pancreatic cancer cell line PaTu8988

Pancreaticadenocarcinomarankssixthofcancer-relateddeathinChina,andittendstobeadvancedatthetimeofpresentation,70%－90%patientsatthetimeofdiagnosishadinoperabledisease.5-Fluorouracil(5FU)iscommonlyusedaloneorwithradiotherapyinthetreatmentofresectablepancreaticcancerandinthepalliationofunresectablepancreaticcancer.Butthecurativeeffectof5FUislimitedbyitssideeffects.Suicidegenetherapyisanewexperimentalformofcancerchemotherapybeingevaluatedinhumantrials.Cytosinedeaminase(CD)isanenzymefoundinava-r…     

Influence of anti-keratin autoantibodies on telomerase activity of squamous cell carcinoma

Objective: To investigate the influence of anti-keratin autoantibodies (AK auto Abs) on telom-erase activity of squamous cell carcinoma cultured in vitro and the mechanisms of the inhibitory effects of AK auto Abs on squamous cell carcinoma. Methods: Influence of AK auto Abs on the proliferation of Tca cells was observed by MTT colorimetry. Telomerase activity of cultured Tca cells and human keratinocytes was determined by telomeric repeat amplication protocol-ELISA (TRAP-ELISA) and polyacrylamide gel elec-trophoresis (PAGE). After being treated with AK auto Abs for 36 h at a concentration of 4, 8, 16 mg/L respectively, the changes of telomerase activity of Tea cells were also detected by TRAP-ELISA and PAGE. Results: MTT colorimetric determination showed that the capacity of proliferation of Tca cells correlated negatively with the concentration of AK auto Abs (r= -0. 74, P<0. 01). TRAP-ELISA and PAGE showed that telomerase activity of Tca cells increased significantly compared to that of cultured human     

Generation of bcl-2 antisense RNA promotes apoptosis of human promyelocytic leukemia cell line HL-60

SupportedbytheNationaINaturalScienceFoundationofChina,No.3957o79ONotonlydoesapoptosisplayakeyroleinhomeostasis,butalsohascloserelationshipwithtumorigenesisandtumorregression,aswellaswiththechemotherapeuticandradiotherapeuticef-fectsontumors[lJ.Justasthecellproliferation,thecellapoptosisisalsomodulatedbymanygenes,includingsomeoncogenesandtumorsup-pressorgenes(p53,bcl-2,c-my,etc.)[2].Forex-ample,thehighexpressionofbcl-2inleukemiacellscanobviouslyinhibittheapoptosisinducedbytheremovalofgrowth…     

Antitumor activity of psoralen on mucoepidermoid carcinoma cell line MEC-1.

Psoralen (PSO) was found cytotoxic against in vitro cultured human mucoepidermoid carcinoma cells of MEC-1 cell line. Its IC50 value was 8.6 micrograms/ml, and relative antitumor activity (RAA) 15. PSO suppressed DNA synthesis, damaged microvilli and cell membrane, and induced degeneration of mitochondria. It inhibited the growth of MEC-1 cells transplanted in nude mice by 79.1% which was as strong as pingyanmycin (PYM 81.4%). The body weight loss of PSO treated tumor-bearing mice was 5%, whereas that of PYM treated ones 13.3% (P < 0.01). PSO may be an effective agent for the treatment of human mucoepidermoid carcinoma.

     

Efects of proglumide, a gastrin receptor antagonist, on human large intestine carcinoma SW480 cell line

Ｓｏｍｅｓｔｕｄｉｅｓ１，２ｉｎｄｉｃａｔｅｄｔｈａｔｇａｓｔｒｉｎｈａｓａｐｒｏｍｏｔｉｎｇｅｆｅｃｔｏｎｔｈｅｇｅｎｅｓｉｓａｎｄｇｒｏｗｔｈｏｆｓｏｍｅｌａｒｇｅｉｎｔｅｓｔｉｎｅｃａｒｃｉｎｏｍａｓ，ａｎｄｐｒｏｇｌｕｍｉｄｅ，ａｇａｓｔｒ...     

Establishment of the human hepatocellular carcinoma cell line HCC－9204 and its characteristics

ＥｓｔａｂｌｉｓｈｍｅｎｔｏｆｔｈｅｈｕｍａｎｈｅｐａｔｏｃｅｌｌｕｌａｒｃａｒｃｉｎｏｍａｃｅｌｌｌｉｎｅＨＣＣ－９２０４ａｎｄｉｔｓｃｈａｒａｃｔｅｒｉｓｔｉｃｓＨｕＣｈｕａｎｍｉｎ；（胡川闽）；ＬｉｕＹａｎｆａｎｇ；（刘彦仿）；ＳｕｉＹａｎｆａ...     

Inhibitory effect of medroxyprogesterone acetate on malignant growth of ovarian cancer 3AO cell line

Ｉｎｈｉｂｉｔｏｒｙｅｆｆｅｃｔｏｆｍｅｄｒｏｘｙｐｒｏｇｅｓｔｅｒｏｎｅａｃｅｔａｔｅｏｎｍａｌｉｇｎａｎｔｇｒｏｗｔｈｏｆｏｖａｒｉａｎｃａｎｃｅｒ　３ＡＯｃｅｌｌｌｉｎｅＪｉｎＺｈｉｊｕｎ（金志军）；ＺｈａｎｇＸｉｙｉｎ（张惜阴）；ＦｅｎｇＹｏ...     

Heparanase antisense oligodeoxynucleotide inhibits angiogenesis and metastasis of human mammary carcinoma cell xenografts in nude mice

Objective： To evaluate the inhibitory effect of heparanase antisense oligodeoxynucleotide （AS-ODN） on the angiogenesis and metastasis of human mammary carcinoma cell xenografts in nude mice. Methods： The AS-ODN complementary to the start codon region of heparanase mRNA and its control, scrambled nonsense oligodeoxynucleotide （NS-ODN） were designed and synthesized. A subcutaneous growth model and an acute hematogenous metastasis model of human mammary carcinoma were established in nude mice and were treated with ODNs. The heparanase expression in tumor was evaluated by RT-PCR and Western blot. The microvessel density （MVD） was measured by immunohistochemistry for factor VS. The tumor volume was calculated and lung metastatic nodules were counted. Results ： The heparanase expression, MVD, tumor volume and lung metastatic nodules in AS-ODN treated group were significantly decreased compared with that in NS-ODN treated group and that in PBS group （P〈0.01）. Conclusion ： Heparanase AS-ODN has significant inhibitory effect on the angiogenesis and metastasis of human mammary carcinoma cell xenografts in nude mice.     

Effect of interleukin-6 on the growth of human lung cancer cell line.

Ｅｆｅｃｔｏｆｉｎｔｅｒｌｅｕｋｉｎ６ｏｎｔｈｅｇｒｏｗｔｈｏｆｈｕｍａｎｌｕｎｇｃａｎｃｅｒｃｅｌｌｉｎｅＦｕＪｉａｎ付坚，ＺｈｅｎｇＪｉｅ郑杰，ＦａｎｇＷｅｉｇａｎｇ方伟岗ａｎｄＷｕＢｉｎｇｑｕａｎ吴秉铨ＤｅｐａｒｔｍｅｎｔｏｆＰａｔｈｏｌｏｇ...     

A study on improvement of expression of human G－CSF cDNA transferred with retroviral double－copy vector

ＡｓｔｕｄｙｏｎｉｍｐｒｏｖｅｍｅｎｔｏｆｅｘｐｒｅｓｓｉｏｎｏｆｈｕｍａｎＧ－ＣＳＦｃＤＮＡｔｒａｎｓｆｅｒｒｅｄｗｉｔｈｒｅｔｒｏｖｉｒａｌｄｏｕｂｌｅ－ｃｏｐｙｖｅｃｔｏｒＧｕｏＢａｏｙｕ（郭葆玉）；ＺｈａｎｇＳｕｙｉｎｇ（张淑英）；ＸｕＨｕｉ...     

Quantitative investigation of invasive effects of laminin-receptor antibodies on human colorectal carcinoma cell lines

Itisoneoftheimportantsubjectstoinvestigatethemechanismofcolorectalcarcinomainvasionandmetastasis,whichmainlyresultinpatients'death.Nowadaysevidencesshowusthatlamininreceptor(I-NR)locatedinplasmamembranecombinedtolaminin,whichisoneofcomponentsofbasementmembrane(BM),isrelevanttocancercellsadhesion,motility,invasionandsignalcommunicationbetweencellandBM.TypetVcollagen,anothercomponentofBMwasdigestedbytypetVcollagenases(IVcol)[1.2),andtheactivitiesoftheseenzymeswerealsorelevanttocancerinvasio…     

Apoptotic effect and mechanisms of AHPN on human skin malignant melanoma cell A375

Objective： To study apoptotic effects of synthetic retinoic acid 6-[3-（1-adamantyl）-4-hydroxyphenyl]-2-naphthalene carboxylic acid（AHPN） on human skin malignant melanoma A375 cells in comparison with the natural ligand all-trans-retinoic acid（ATRA） in vitro and the mechanisms related to the actions of AHPN. Methods：MTT assay was used to determine the anti-proliferative effects of AHPN and ATRA on A375 cells. Flow cytometry was performed to investigate the influence of AHPN and ATRA on cell cycle and cell apoptosis. In addition, transfection and luciferase activity assays were employed to explore the mechanisms of how AHPN executes its proapoptotic function. Results：Firstly, AHPN promoted apoptosis and GI arrest in A375 cells compared with ATRA. Secondly, the activity of NF-K B in A375 cells treated with AHPN increased 2-3 times compared with solvent DMSO treatment. Conclusion：AHPN, in comparison with ATRA, is a more effective alternative for therapy of malignant melanoma. The potentially proapoptotic function of AHPN requires activation of NF-K B.     

The effects of genistein on TGF-β1-induced the invasion and metastasis in human pancreatic cancer cell line Panc-1

Pancreatic cancer is a devastating disease with the worst mortality rate. Therefore, a rational strategy for future drug development is critical. Genistein is a small, biologically active flavonoid that is found in high amounts in soy. This important compound possesses a wide variety of biological activities, but it is best known for its ability to inhibit cancer progression. In the current study, we found that genistein can inhibit effectively TGF-β1-induced the invasion and metastasis in Panc-1 by Transwell assay, which is through regulating the mRNA and protein expression of uPA and MMP2, but not MMP9 by RT-PCR / Weston blot, and is positively correlated with the concentration of genistein. At the same time, genistein also could improve the progress of Epithelial-Mesenchymal Transition (EMT) via morphology observation using light microscope / TEM, which is mediated by the down-regulation of E-cadherin and the up-regulation of vimentin. TGF-β1 mediates EMT process via numerous intracellular signal transduction pathways. The potential molecular mechanisms are all or partly through Smad4-dependent and -independent pathways (p38 MAPK) to regulate the antitumor effect of genistein.