Identification of up-regulated and down-regulated cis-natural antisense transcripts in the human B lymphoblastic cell line IM-9 after X-ray irradiation

Chiba, Mitsuru; Miura, Tomisato; Kasai, Kosuke; Monzen, Satoru; Kashiwakura, Ikuo; Yasue, Hiroshi; Nakamura, Toshiya

doi:10.3892/mmr.2012.787

Cited by 7 publications

(10 citation statements)

References 15 publications

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“…In addition, we detected the natural antisense RNAs of MDM2 and CDKN1A genes in human cell lines in a previous study (23). Therefore, in order to examine whether natural antisense RNAs are present within exosomes, the natural antisense RNAs of LRRC24, MDM2 and CDKN1A genes were examined by strand-specific RT-PCR as described in Materials and methods.…”

Section: Resultsmentioning

confidence: 97%

“…Kohno et al discovered that the natural antisense RNA of LRRC24 gene was upregulated in the human CRC tissues, compared with normal colon tissues (19). Furthermore, we detected the natural antisense RNAs of MDM2 and CDKN1A genes in human B lymphoblastic cells in a previous study, and showed that their natural antisense RNAs were in the cytoplasm (23). In the present study, it was clarified that their intracellular natural antisense RNAs were enclosed within exosomes.…”

Section: Discussionmentioning

confidence: 94%

“…To synthesize cDNAs derived from natural antisense RNAs, reverse transcription reactions were performed using exosomal RNAs (1 ng), the forward primers shown in Table III, and an AMV Reverse Transcriptase (Promega, Madison, WI) (19,23,24). PCRs were performed using cDNAs derived from natural antisense RNAs, a Power SYBR Green Master mix (Life Technologies), the primer pairs described in Table III, and the StepOne Plus Real-Time PCR system (Life Technologies) under the following conditions: 10 min at 95°C, followed by 40 cycles each of 95°C for 15 sec and 60°C for 60 sec.…”

Section: Methodsmentioning

confidence: 99%

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Exosomes secreted from human colorectal cancer cell lines contain mRNAs, microRNAs and natural antisense RNAs, that can transfer into the human hepatoma HepG2 and lung cancer A549 cell lines

2012

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Exosomes are microvesicles that are released from various cells into the extracellular space. It has been reported that the components within exosomes vary according to the type of secreted cell. In the present study, we investigated the tetraspanin family proteins CD63, CD9 and CD81 as useful collection markers of exosomes derived from the three colorectal cancer (CRC) cell lines HCT-15, SW480 and WiDr. In addition, we aimed to detect the mRNAs, microRNAs and natural antisense RNAs within the exosomes secreted from the three CRC cell lines. Furthermore, we examined whether exosomes containing their RNAs were transferred into the hepatoma cell line HepG2 and lung cancer cell line A549. CD81 was detected in exosomes secreted from the three CRC cell lines. This result indicates that CD81 can be a collection marker of exosomes derived from the three CRC cell lines. When the RNA species within exosomes derived from the three CRC cell lines were examined, the mRNAs of housekeeping genes such as ACTB and GAPDH, the microRNAs such as miR-21, miR-192 and miR-221, and the natural antisense RNAs of LRRC24, MDM2 and CDKN1A genes, were detected. We discovered their natural antisense RNAs within exosomes for the first time in the present study. Furthermore, PKH67-labeled exosomes derived from the CRC cell lines were taken up into HepG2 and A549 cells. These findings indicate that the intracellular RNAs enclosed within exosomes are secreted to the outside, and exosomes derived from the CRC cell lines are transferred into HepG2 and A549 cells. In conclusion, we reveal that exosomes derived from the CRC cell lines contain mRNAs, microRNAs and natural antisense RNAs, and can be delivered into HepG2 and A549 cells. These findings indicate that exosomal RNAs can shuttle between cells, and may be involved in the regulation of gene expression in recipient cells.

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Section: Resultsmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

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Exosomes secreted from human colorectal cancer cell lines contain mRNAs, microRNAs and natural antisense RNAs, that can transfer into the human hepatoma HepG2 and lung cancer A549 cell lines

2012

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“…Lymphoblastoid cell lines (LCL), for example, have been used to investigate differences in the function of pathways involved in DNA repair and apoptosis (Veldwijk et al 2011, Chiba et al 2012). They have also been used to identify genetic determinants of pathways involved in the cytotoxic response to IR (Shukla and Dolan 2005, Hartford and Dolan 2007, Niu et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Radiation-induced apoptosis varies among individuals and is modified by sex and age

Applebaum

Skol

Bond

et al. 2014

International Journal of Radiation Biology

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Purpose Although there are considerable data on mechanisms of radiation-induced apoptosis in vitro and in animal models, little is known about functional variation in these pathways in humans. We sought to develop a tractable system to evaluate this. Materials and methods Peripheral blood mononuclear cells were isolated from 90 healthy volunteers, divided into two aliquots, one irradiated with a 5 Gy dose and the other sham-treated (0 Gy), and assessed for damage-induced apoptosis after 24 hours. To investigate reproducibility, ten individuals spanning the entire radiation-induced apoptotic range were tested three times each, with 3–6 months between replicates. Results We observed surprising heterogeneity in apoptosis among individuals, ranging from 21–62%. Biological replicates from a single individual, however, were completely concordant, suggesting the variability observed across individuals is not the result of stochastic or short-term effects. We found significantly higher radiation-induced apoptosis in males than in females (Mean: 41.0% vs. 30.7%; p < 3.5 × 10−7). Moreover, advancing age was associated with decreasing radiation-induced apoptosis in males (p = 0.01) but not females (p = 0.82).a Conclusions Our results provide evidence that the function of cellular pathways crucial for stress-induced apoptosis varies by sex and could decline with age in humans.

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“…Natural antisense transcripts (NATs) are known as one of gene expression regulators in eukaryotes [7]. In the past several years, the expression of NATs in mammals is reported [8,9]. In addition, investigation that the differential expressions of several NATs are induced during the process of liver development is made [10].…”

Section: Introductionmentioning

confidence: 99%

The expressions of Caveolin-2 mRNAs, natural antisense transcripts and proteins in mouse liver development

Chiba¹

2018

biomedicalresearch

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Caveolin-2 (Cav2) has been shown to be a major component of the inner surface of caveolae and be involved in lipid metabolism, development and cancer. Natural Antisense Transcripts (NATs) are transcribed from a large number of genes in various species, including humans and mice. In the present study, we investigated the expressions and localization of Cav2 mRNAs, NATs and proteins in the mouse liver at various developmental stages. The expressions of Cav2 mRNAs and proteins in the liver were higher at the new-born stage (NB) than at days 14, 17, and 19 of the embryonic stage (E) and at day 3 after birth. The expressions of Cav2 NATs were lower than that of Cav2 mRNAs at each stage. On tissues, the expressions of Cav2 mRNAs and NATs were uniformly low in the liver at E14. The expressions of Cav2 mRNAs were predominantly strong in the hepatocytes at NB, whereas the expressions of Cav2 NATs were low in the hepatocytes at NB but strongly detected in specific cells. In conclusions, Cav2 mRNAs and NATs did not co-localized in the liver at NB. These results suggest that Cav2 NATs may be not involved in the regulation of Cav2 mRNA expressions at NB.

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Identification of up-regulated and down-regulated cis-natural antisense transcripts in the human B lymphoblastic cell line IM-9 after X-ray irradiation

Cited by 7 publications

References 15 publications

Exosomes secreted from human colorectal cancer cell lines contain mRNAs, microRNAs and natural antisense RNAs, that can transfer into the human hepatoma HepG2 and lung cancer A549 cell lines

Exosomes secreted from human colorectal cancer cell lines contain mRNAs, microRNAs and natural antisense RNAs, that can transfer into the human hepatoma HepG2 and lung cancer A549 cell lines

Radiation-induced apoptosis varies among individuals and is modified by sex and age

The expressions of Caveolin-2 mRNAs, natural antisense transcripts and proteins in mouse liver development

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