title: Chemosensitization of HepG2 cells by suppression of NF-κB/p65 gene transcription with specific-siRNA.
abstract: OBJECTIVE
  To investigate small interfering RNA (siRNA)-mediated inhibition of nuclear factor-kappa B (NF-κB) activation and multidrug-resistant (MDR) phenotype formation in human HepG2 cells.
  
  
  METHODS
  Total RNA was extracted from human HepG2 or LO2 cells. NF-κB/p65 mRNA was amplified by nested reverse transcription polymerase chain reaction and confirmed by sequencing. NF-κB/p65 was analyzed by immunohistochemistry. Specific-siRNA was transfected to HepG2 cells to knock down NF-κB/p65 expression. The effects on cell proliferation, survival, and apoptosis were assessed, and the level of NF-κB/p65 or P-glycoprotein (P-gp) was quantitatively analyzed by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  HepG2 cells express NF-κB/p65 and express relatively less phosphorylated p65 (P-p65) and little P-gp. After treatment of HepG2 cells with different doses of doxorubicin, the expression of NF-κB/p65, P-p65, and especially P-gp were dose-dependently upregulated. After HepG2 cells were transfected with NF-κB/p65 siRNA (100 nmol/L), the expression of NF-κB/p65, P-p65, and P-gp were downregulated significantly and dose-dependently. The viability of HepG2 cells was decreased to 23% in the combination NF-κB/p65 siRNA (100 nmol/L) and doxorubicin (0.5 μmol/L) group and 47% in the doxorubicin (0.5 μmol/L) group (t = 7.043, P < 0.001).
  
  
  CONCLUSIONS
  Knockdown of NF-κB/p65 with siRNA is an effective strategy for inhibiting HepG2 cell growth by downregulating P-gp expression associated chemosensitization and apoptosis induction.
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Si-Ye
authors.initials: SY
authors.lastname: Wang
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Min
authors.initials: M
authors.lastname: Yao
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Wen-Li
authors.initials: WL
authors.lastname: Sai
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Jun-Ling
authors.initials: JL
authors.lastname: Yang
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Cai
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Wen-Jie
authors.initials: WJ
authors.lastname: Zheng
authors.affiliation: Yun Shi, Si-Ye Wang, Min Yao, Wen-Li Sai, Wei Wu, Jun-Ling Yang, Wen-Jie Zheng, Deng-Fu Yao, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
authors.forename: Deng-Fu
authors.initials: DF
authors.lastname: Yao
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics / Q000378:metabolism; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology*; D018432:Drug Resistance, Multiple / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D008297:Male; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D000067492:RNAi Therapeutics; D013997:Time Factors; D051996:Transcription Factor RelA / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D000903:Antibiotics, Antineoplastic; C498143:RELA protein, human; D034741:RNA, Small Interfering; D051996:Transcription Factor RelA; D004317:Doxorubicin
keywords: Chemosensitization; Hepatocellular carcinoma; Multidrug-resistant; Nuclear factor-κB; P-glycoprotein; Small interference RNA