慢病毒介导RNA干扰HES1鼻咽癌稳定细胞株的建立与鉴定
目的建立稳定干扰Split多毛增强子1（HES1）的鼻咽癌（NPC）细胞株,为进一步探讨HES1对NPC的影响奠定基础。方法将293T细胞接种6孔板后随机分为A、B、C、D、E组,分别瞬时转染慢病毒质粒shHES1-a、shHES1-b、shHES1-c、shHES1-d及空载质粒shSCR,分别用qRT-PCR和Western blot法检测HES1 mRNA及蛋白,筛选出最有效干扰质粒;将最有效干扰质粒或空载质粒与慢病毒包装质粒共转染293T细胞,获得含最有效干扰质粒或空载质粒的病毒上清（LV-shHES1/LV-shSCR）。将NPC细胞CNE2和S18随机分为观察组和对照组,分别将LV...
Saved in:
| Published in | 山东医药 Vol. 57; no. 18; pp. 9 - 11 |
|---|---|
| Main Author | |
| Format | Magazine Article |
| Language | Chinese |
| Published |
南方医科大学肿瘤研究所,广州,510515
2017
|
| Subjects | |
| Online Access | Get full text |
| ISSN | 1002-266X |
| DOI | 10.3969/j.issn.1002-266X.2017.18.003 |
Cover
| Abstract | 目的建立稳定干扰Split多毛增强子1（HES1）的鼻咽癌（NPC）细胞株,为进一步探讨HES1对NPC的影响奠定基础。方法将293T细胞接种6孔板后随机分为A、B、C、D、E组,分别瞬时转染慢病毒质粒shHES1-a、shHES1-b、shHES1-c、shHES1-d及空载质粒shSCR,分别用qRT-PCR和Western blot法检测HES1 mRNA及蛋白,筛选出最有效干扰质粒;将最有效干扰质粒或空载质粒与慢病毒包装质粒共转染293T细胞,获得含最有效干扰质粒或空载质粒的病毒上清（LV-shHES1/LV-shSCR）。将NPC细胞CNE2和S18随机分为观察组和对照组,分别将LVshHES1、LV-shSCR以浓度梯度方式感染细胞,分别采用qRT-PCR和Western blot法检测HES1 mRNA及蛋白。结果C组293T细胞中HES1 mRNA及蛋白相对表达量均低于其余各组（P均〈0.05）。与对照组比较,观察组CNE2和S18细胞中HES1 mRNA及蛋白相对表达量均降低（P均〈0.05）。结论成功建立稳定干扰HES1的NPC细胞株。 |
|---|---|
| AbstractList | 目的建立稳定干扰Split多毛增强子1（HES1）的鼻咽癌（NPC）细胞株,为进一步探讨HES1对NPC的影响奠定基础。方法将293T细胞接种6孔板后随机分为A、B、C、D、E组,分别瞬时转染慢病毒质粒shHES1-a、shHES1-b、shHES1-c、shHES1-d及空载质粒shSCR,分别用qRT-PCR和Western blot法检测HES1 mRNA及蛋白,筛选出最有效干扰质粒;将最有效干扰质粒或空载质粒与慢病毒包装质粒共转染293T细胞,获得含最有效干扰质粒或空载质粒的病毒上清（LV-shHES1/LV-shSCR）。将NPC细胞CNE2和S18随机分为观察组和对照组,分别将LVshHES1、LV-shSCR以浓度梯度方式感染细胞,分别采用qRT-PCR和Western blot法检测HES1 mRNA及蛋白。结果C组293T细胞中HES1 mRNA及蛋白相对表达量均低于其余各组（P均〈0.05）。与对照组比较,观察组CNE2和S18细胞中HES1 mRNA及蛋白相对表达量均降低（P均〈0.05）。结论成功建立稳定干扰HES1的NPC细胞株。 R739.62; 目的 建立稳定干扰Split多毛增强子1(HES1)的鼻咽癌(NPC)细胞株,为进一步探讨HES1对NPC的影响奠定基础.方法 将293T细胞接种6孔板后随机分为A、B、C、D、E组,分别瞬时转染慢病毒质粒shHES1-a、shHES1-b、shHES1-c、shHES1-d及空载质粒shSCR,分别用qRT-PCR和Western blot法检测HES1 mRNA及蛋白,筛选出最有效干扰质粒;将最有效干扰质粒或空载质粒与慢病毒包装质粒共转染293T细胞,获得含最有效干扰质粒或空载质粒的病毒上清(LV-shHES1/LV-shSCR).将NPC细胞CNE2和S18随机分为观察组和对照组,分别将LV-shHES1、LV-shSCR以浓度梯度方式感染细胞,分别采用qRT-PCR和Western blot法检测HES1 mRNA及蛋白.结果 C组293T细胞中HES1 mRNA及蛋白相对表达量均低于其余各组(P均<0.05).与对照组比较,观察组CNE2和S18细胞中HES1 mRNA及蛋白相对表达量均降低(P均<0.05).结论 成功建立稳定干扰HES1的NPC细胞株. |
| Abstract_FL | Objective To establish a nasopharyngeal carcinoma (NPC) cell line for silencing hairy and enhancer of split homolog-1 (HES1), and it will lay a foundation for further study on the effect of HES1 on NPC.Methods cell 293T inoculated on a 6-well plate were divided into groups A, B, C, D and E, which were instantaneously transfected with the recombinant lentiviruses (shHES1-a, shHES1-b, shHES1-c, shHES1-d and empty vector shSCR).The expression of HES1 was analyzed by qRT-PCR and Western blotting to identify the LV-shHES1 vector that had the highest gene knockdown efficiency.cell 293T were co-transfected with the most effective interference plasmid or no-load plasmid and lentiviral packaging plasmid to obtain the virus supernatant (LV-shHes1/LV-shSCR) containing the most effective interference plasmid or no-load plasmid.NPC cells CNE2 and S18 were randomly divided into the observation group and control group.LV-shHes1 and LV-shSCR were used to infect the cells in a concentration gradient manner, and the mRNA and protein of HES1 was detected by qRT-PCR and Western blot.Results The relative expression of HES1 mRNA and protein in 293T cells in the group C was lower than that in the other groups, and the difference was statistically significant (all P<0.05).Therefore, the shHES1-c interference efficiency was the best.Compared with the control group, the relative expression of HES1 mRNA and protein in both CNE2 and S18 cells of the observation group was decreased (all P<0.05).Conclusion NPC cell line for silencing HES1 gene by RNA interference was successfully established. |
| Author | 程玉霜 王慧艳 肖东 |
| AuthorAffiliation | 南方医科大学肿瘤研究所,广州510515 |
| AuthorAffiliation_xml | – name: 南方医科大学肿瘤研究所,广州,510515 |
| Author_FL | CHENG Yushuang WANG Huiyan XIAO Dong |
| Author_FL_xml | – sequence: 1 fullname: CHENG Yushuang – sequence: 2 fullname: WANG Huiyan – sequence: 3 fullname: XIAO Dong |
| Author_xml | – sequence: 1 fullname: 程玉霜 王慧艳 肖东 |
| BookMark | eNo9jz9Lw0AYh2-oYFv9EuLgknh3udwlYymtFYqCdnALl0vSP-hVG0Q6a4VScegidhBxEgUr6qCmfpsmsd_Ck4rTy_u-D78fTw5kZFv6AKwiqBs2tddbejMMpY4gxBqmdE_HEDEdWTqERgZk_--LIBeGLQgJRszOgnLSu0uvesl4OI0G8Xiys1WIP16S_nOltItmkygefqXXF-n9a_w0SqPz79Ob5PYhHZ3F0Wf6OJi-X876b-q1BBYCvh_6y38zD2rlUq1Y0arbG5vFQlUTFBKNBIFgLmeuRTjlwhceJ4QHLjXUapKAUGGaxIWuhyB2henbAcbC9jC2PIY8y8iDtXnsCZcBl3Wn1T7uSFXohA0uvW73VxpZyk6hK3NUNNqyftRU8GGnecA7XYcy5Y4xM4wfYXB0VA |
| ClassificationCodes | R739.62 |
| ContentType | Magazine Article |
| Copyright | Copyright © Wanfang Data Co. Ltd. All Rights Reserved. |
| Copyright_xml | – notice: Copyright © Wanfang Data Co. Ltd. All Rights Reserved. |
| DBID | 2RA 92L CQIGP W91 ~WA 2B. 4A8 92I 93N PSX TCJ |
| DOI | 10.3969/j.issn.1002-266X.2017.18.003 |
| DatabaseName | 维普期刊资源整合服务平台 中文科技期刊数据库-CALIS站点 维普中文期刊数据库 中文科技期刊数据库-医药卫生 中文科技期刊数据库- 镜像站点 Wanfang Data Journals - Hong Kong WANFANG Data Centre Wanfang Data Journals 万方数据期刊 - 香港版 China Online Journals (COJ) China Online Journals (COJ) |
| DatabaseTitleList | |
| DeliveryMethod | fulltext_linktorsrc |
| DocumentTitleAlternate | Establishment and identification of NPC cell line for silencing HES1 gene by RNA interference |
| DocumentTitle_FL | Establishment and identification of NPC cell line for silencing HES1 gene by RNA interference |
| EndPage | 11 |
| ExternalDocumentID | shandyy201718004 672172273 |
| GrantInformation_xml | – fundername: 国家自然科学基金资助项目; 广东省自然科学基金资助项目 funderid: (81172587,81372896,81672689); (2014A030313294) |
| GroupedDBID | -05 2B. 2C~ 2RA 5XA 5XF 92F 92I 92L ABDBF ACGFS ALMA_UNASSIGNED_HOLDINGS CCEZO CIEJG CQIGP CW9 EOJEC OBODZ TCJ TGQ U1G U5O W91 ~WA 4A8 93N ABJNI ACUHS PSX |
| ID | FETCH-LOGICAL-c604-4ffc7ba7b84a6acecda44afb63a6a54f46c554b0bd102bc5e9f22c9d228d71d83 |
| ISSN | 1002-266X |
| IngestDate | Thu May 29 04:06:27 EDT 2025 Wed Feb 14 10:03:16 EST 2024 |
| IsPeerReviewed | false |
| IsScholarly | true |
| Issue | 18 |
| Keywords | RNA干扰 nasopharyngeal carcinoma hairy and enhancer of split homolog-1 Split多毛增强子1 RNA interference NPC cells 慢病毒载体 鼻咽癌 NPC细胞株 lentivirus expression vector |
| Language | Chinese |
| LinkModel | OpenURL |
| MergedId | FETCHMERGED-LOGICAL-c604-4ffc7ba7b84a6acecda44afb63a6a54f46c554b0bd102bc5e9f22c9d228d71d83 |
| Notes | 37-1156/R nasopharyngeal carcinoma; hairy and enhancer of split homolog-1; RNA interference; lentivirus expression vector; NPC cells Objective To establish a nasopharyngeal carcinoma（ NPC） cell line for silencing hairy and enhancer of split homolog-1（ HES1）,and it will lay a foundation for further study on the effect of HES1 on NPC. Methods cell 293 T inoculated on a 6-well plate were divided into groups A,B,C,D and E,which were instantaneously transfected with the recombinant lentiviruses（ shHES1-a,shHES1-b,shHES1-c,shHES1-d and empty vector shSCR）. The expression of HES1 was analyzed by qRT-PCR and Western blotting to identify the LV-shHES1 vector that had the highest gene knockdown efficiency. cell 293 T were co-transfected with the most effective interference plasmid or no-load plasmid and lentiviral packaging plasmid to obtain the virus supernatant（ LV-shHes1/LV-shSCR） containing the most effective interference plasmid or no-load plasmid. NPC cells CNE2 and S18 were randomly divided into the observation gr |
| PageCount | 3 |
| ParticipantIDs | wanfang_journals_shandyy201718004 chongqing_primary_672172273 |
| PublicationCentury | 2000 |
| PublicationDate | 2017 |
| PublicationDateYYYYMMDD | 2017-01-01 |
| PublicationDate_xml | – year: 2017 text: 2017 |
| PublicationDecade | 2010 |
| PublicationTitle | 山东医药 |
| PublicationTitleAlternate | Shandong Medical Journal |
| PublicationTitle_FL | Shandong Medical Journal |
| PublicationYear | 2017 |
| Publisher | 南方医科大学肿瘤研究所,广州,510515 |
| Publisher_xml | – name: 南方医科大学肿瘤研究所,广州,510515 |
| SSID | ssj0042179 ssib001104105 ssib017477535 ssib051371736 ssib002262813 |
| Score | 1.3331076 |
| Snippet | 目的建立稳定干扰Split多毛增强子1（HES1）的鼻咽癌（NPC）细胞株,为进一步探讨HES1对NPC的影响奠定基础。方法将293T细胞接种6孔板后随机分为A、B、C、D、E组,分别瞬时转染慢病... R739.62; 目的 建立稳定干扰Split多毛增强子1(HES1)的鼻咽癌(NPC)细胞株,为进一步探讨HES1对NPC的影响奠定基础.方法 将293T细胞接种6孔板后随机分为A、B、C、D、E组,分别瞬时... |
| SourceID | wanfang chongqing |
| SourceType | Aggregation Database Publisher |
| StartPage | 9 |
| SubjectTerms | NPC细胞株 RNA干扰 Split多毛增强子1 慢病毒载体 鼻咽癌 |
| Title | 慢病毒介导RNA干扰HES1鼻咽癌稳定细胞株的建立与鉴定 |
| URI | http://lib.cqvip.com/qk/92062X/201718/672172273.html https://d.wanfangdata.com.cn/periodical/shandyy201718004 |
| Volume | 57 |
| hasFullText | 1 |
| inHoldings | 1 |
| isFullTextHit | |
| isPrint | |
| link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnR3LahRBcIgJiCcVFWNUIqSPG-fR04_j9OwsQTAHXSG3ZXoeyWmjJjkkV40QIh5yEXMQ8SQKRtSDuvFvsrvmL6zqmZ0dgkgUlqGmurqqtmq7q3q2usey5lJXZLmG1UnqpbRBc5gHJc3cBtVcy9S3Y5nh5uS7i2zhAb2z5C9NnNG1qqWNdT2fbP1xX8n_eBVw4FfcJfsPnq2YAgJg8C9cwcNwPZWPScSI8EngkogTyREGTNAiEjCUKEWEIpGPGBXeWwwQVpIo13QEwF6I7jskAihEamiGnqpp2EkiQgQCQZRnuEREBohBvoxEggiPyMiItElgmoBAUCNGEWUwQaEEaAP0EcpCwbRiWM-OTUcHPwW9DBEjCt2gexO-yegXUqqGvDkyBq7AG3rIsE6CLWpkJ264gHhvTAIIl0hWSaw_BnH4iccgRp0mmho4Qicw5lhBjgKkY74ZNcIAAJVZKUS1jIUEtiKxTWQBgEKs9AhWYoYjN7UMwNHGbogparEltYwhGGTc4gWNVZApTuEeDSZRCxmylnsUcedkVPMkkyaqIfv5ij3WJfJ5B2uBvXE0r2os1_Cfl81NJHKEOTJ3yuWQX05aU4FqqtY4aYYVuVOb9SEhd8U4yfMdDys12Ci_obCIlaaGoNTjrDVXKnn7byri4SUrq93lR5CSmR1y3TzuLteSufYF6_zoVPXZoBhTF62JrZVLVmuw_Xb4cntwsHfU2-0fHMKI6X__PNj5hKPk-LDX3_s5fPV8-O5L_-P-sPfs15PXgzfvh_tP-70fww-7R99eHO98habLVrsVtcOFRvmekUbCbJyg8oTrmGtBYxYnWZLGlMa5Zh7c-jSnLIGcW9s6hWRcJ34mc9dNZOq6IuVOKrwr1mR3tZtdtWa1TFydUy_2bUpl7IsUbmw7zpLcS5mW09ZMZYPOw-I4mQ7j-JI4WEZMW7dKq3TKSWatc9KL105BM2OdG4-Q69bk-uON7Aakzuv6Zun736U1mzM |
| linkProvider | EBSCOhost |
| openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=%E6%85%A2%E7%97%85%E6%AF%92%E4%BB%8B%E5%AF%BCRNA%E5%B9%B2%E6%89%B0HES1%E9%BC%BB%E5%92%BD%E7%99%8C%E7%A8%B3%E5%AE%9A%E7%BB%86%E8%83%9E%E6%A0%AA%E7%9A%84%E5%BB%BA%E7%AB%8B%E4%B8%8E%E9%89%B4%E5%AE%9A&rft.jtitle=%E5%B1%B1%E4%B8%9C%E5%8C%BB%E8%8D%AF&rft.au=%E7%A8%8B%E7%8E%89%E9%9C%9C&rft.au=%E7%8E%8B%E6%85%A7%E8%89%B3&rft.au=%E8%82%96%E4%B8%9C&rft.date=2017-01-01&rft.pub=%E5%8D%97%E6%96%B9%E5%8C%BB%E7%A7%91%E5%A4%A7%E5%AD%A6%E8%82%BF%E7%98%A4%E7%A0%94%E7%A9%B6%E6%89%80%2C%E5%B9%BF%E5%B7%9E%2C510515&rft.issn=1002-266X&rft.volume=57&rft.issue=18&rft.spage=9&rft.epage=11&rft_id=info:doi/10.3969%2Fj.issn.1002-266X.2017.18.003&rft.externalDocID=shandyy201718004 |
| thumbnail_s | http://utb.summon.serialssolutions.com/2.0.0/image/custom?url=http%3A%2F%2Fimage.cqvip.com%2Fvip1000%2Fqk%2F92062X%2F92062X.jpg http://utb.summon.serialssolutions.com/2.0.0/image/custom?url=http%3A%2F%2Fwww.wanfangdata.com.cn%2Fimages%2FPeriodicalImages%2Fshandyy%2Fshandyy.jpg |