Combined RNAi targeting human Stat3 and ADAM9 as gene therapy for non-small cell lung cancer
Authors: Fangchao Gong
Youbin Cui
Zhihong Li
Liang Chang
Hongfei Cai
Journal: Oncology Letters (2016)
Discipline: CLINICAL MEDICINE
Keywords: RNA silencing; disintegrin and metalloproteinase 9; non-small cell lung cancer; signal transducer and activator of transcription 3.
Affiliations: Department of Thoracic Surgery, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China.

Abstract:  Previous studies have demonstrated that human signal transducer and activator of transcription 3 (Stat3) and disintegrin and metalloproteinase 9 (ADAM9) are promising targets for RNA interference (RNAi)-based gene therapy for human non-small cell lung cancer (NSCLC). Thus, in the present study, the recombinant lentiviral (Lv) small hairpin (sh)RNA expression plasmids Lv/sh-Stat3 and Lv/sh-ADAM9, which targeted Stat3 and ADAM9, respectively, were constructed and subsequently infected into the A549 human NSCLC cell line. Cell proliferation, migration, invasion and apoptosis were determined in vitro in A549 cells following treatment with Lv/sh-Stat3 or Lv/sh-ADAM9 alone or in combination. In addition, the combined effect of Lv/sh-Stat3 and Lv/sh-ADAM9 gene therapy was evaluated in vivo using A549 xenograft models in nude mice. The in vitro experiments demonstrated that A549 cells treated with a combination of Lv/sh-Stat3 and Lv/sh-ADAM9 exhibited a significant additive effect in their cell proliferation, migration, invasion and apoptosis abilities, compared with A549 cells treated with Lv/sh-Stat3 or Lv/sh-ADAM9 alone. The in vivo experiments conducted in A549 xenograft tumor mouse models revealed that the combined treatment with Lv/sh-Stat3 and Lv/sh-ADAM9 exerted an additive effect on tumor growth inhibition, compared with the treatment with Lv/sh-Stat3 or Lv/sh-ADAM9 alone. These results suggested that combined RNAi gene therapy targeting human Stat3 and ADAM9 may be a novel and promising strategy for the treatment of NSCLC. 

Retracted: No

PubPeer comments:
 Fig 4C.  Western blot analysis of the protein expression levels of MMP-2 and MMP-9 following infection with Lv sh-Stat3 and Lv sh-ADAM9, alone or in combination . ![file] The loading control has appeared in a number of other papers. E.g. Fig 5D from [Han et al (2016)] . ![] The substantive lanes also make other appearances. Fig 5B from [Wen et al (2015)] : ![] *[above]* Fig 5B from [Dong et al (2016)] .  Western blot analysis of CXCL12 protein expression in primary PTC tissues (T) and the matched adjacent normal tissues (ANT) from the same patient. *[below]* Fig 4C from Wen et al (2015).  Western blotting was used to detect LP6 protein expression in TPC-1 cells after transfected with miR-126 mimic or miR-Ctrl. u03b2-actin was the internal control. ![] , 
 Fig 1C.  Western blot analysis of Stat3 and ADAM9 in A549 cells infected with Lv sh-Stat3 and Lv sh-ADAM9, alone or in combination, using specific antibodies. ![file] After a horizontal flip, the ADAM-9 lane displays a high similarity to ADAM-10 from Fig 5D of [Liu et al (2016)] . Red lane seems to have strayed over from Fig 3E of [Liu et al (2015)] . ![file] The loading control makes numerous other appearances, e.g. in Fig 5D from [Wu et al (2015)] . ![] , 
 *[right]* Fig 5A.  Images of tumor tissue from the different treatment groups following sacrifice on day 21 of the treatment. *[left]* Fig 5A from [Zhang et al (2015)] .  Images of tumor tissue with different plasmid treatments collected after sacrification at day 21 . ![file] Could the authors of either paper comment on the appearance of the same tumors in both? UPDATE: Fig 5A from [Chang et al (2015)] .  Tumor images of mice 21 days following treatment with Lv sh-ADAM9 or Lv sh-NC. ![file] ,